15th World Congress on Structural Biology March 18-19,2019 Paris, France

Biography:

Magnus S. Magnusson, Research Professor, founder and director of the Human Behavior Laboratory, University of Iceland. PhD in 1983, University of Copenhagen. Author of the T-pattern model and detection software THEMETM (PatternVision.com), focused on real-time organization of behavior. Co-directed DNA analysis. Numerous papers and talks/keynotes in ethology, neuroscience, mathematics, religion, proteomics, mass spectrometry and nanoscience. Deputy Director 1983-1988, in Museum of Mankind, National Museum of Natural History, Paris. Repeatedly invited temporary Professor at the University of Paris, V, VIII and XIII.  In collaboration between now 32 universities initiated 1995 at University of Paris V, Sorbonne, based on “Magnusson’s analytical model”.

Abstract:

This talk presents a self-similar pattern type called T-pattern, a kind of statistical pseudo fractal recurring with significant translation symmetry on a single discrete dimension (now with a specialized detection algorithm and software THEME for Windows (see patternvision.com), which has allowed the discovery of numerous and complex interaction patterns in many kinds of human and animal interactions as well as in neuronal interactions within living brains. T-patterns have also been detected in interactions between robots and humans and seem characteristic for the structure of DNA and text. A definition of T-patterns is presented as well as the essentials of the current detection algorithms and examples. The potential importance of T-patterns is finally illustrated through a comparison between human mass societies and the mass societies of proteins within biological cells (sometimes called “Cell City”), where self-similarity of organization evolved over billions of years is striking from nano to human scales based on self-similar T-patterns, but appearing suddenly among large-brain animals in humans only and based on massively copied standardized T-patterned letter strings such as holy, legal and scientific texts. The invention of writing and thus a durable external T-patterned memory only a few thousand years ago -- a biological eye-blink -- allowing socio-cultural memory to become largely external to brains and the rise of the only large-brained mass-societies and advanced science and technology. The analogy and self-similarity is striking with the invention of DNA by the RNA world countless millions of years ago.

Biography:

Shuntaro Hara received his M.S. and Ph.D. degrees from the University of Tokyo and started his research career as a postdoctoral fellow at the University of Tokyo. After postdoctoral training, Dr. Hara has worked as a research associate at the National Cardiovascular Center Research Institute (Osaka, Japan), as a visiting fellow at University College London, and as an assistant professor at Kitasato University (Tokyo, Japan), and then became a professor at the School of Pharmacy, Showa University in 2009. He has published more than 100 papers in reputed journals.

Abstract:

Acyl coenzyme A synthetase long-chain family members (ACSLs) are a family of enzymes that convert long-chain free fatty acids into their acyl-CoAs. Among ACSL isozymes, ACSL4 has been hypothesized to modulate the metabolic fates of polyunsaturated fatty acids including arachidonic acid. In the present study, to investigate the enzymatic and protein characteristics of ACSL4, the cDNA for human ACSL4 was cloned from human epithelial colorectal adenocarcinoma Caco-2 cells and then recombinant ACSL4 enzyme containing a C-terminal His-tag was expressed in Spodoptera frugiperda 9 (Sf9) cells using the baculovirus expression system. ACSL4 enzyme activity was detected in 10,000 x g supernatants of ACSL4-expressing Sf9 cell lysates and then partially purified by nickel affinity column chromatography. We further investigated the substrate specificity of recombinant human ACSL4 by LC/MS and found that ACSL4 enzyme preferred various kinds of polyunsaturated fatty acid including docosahexaenoic acid, docosapentanoic acid, eisopentaenoic acid, and dihomo-γ-linolenic acid, as well as arachidonic acid as a substrate. On the other hand, oleic acid, linoleic acid and linolenic acid were poor substrates, although these fatty acids contain unsaturated bonds. These results confirmed the importance of ACSL4 in maintenance of membrane phospholipid bearing polyunsaturated fatty acid.

Biography:

Ina  Buchholz  has  studied  Biochemistry  at  the  University  of  Greifswald,  with  main  focus  on Analytical  Biochemistry  and  Biophysics.  Currently,  she  is  a  PhD  student  in  the  group  of  Prof. Mihaela   Delcea   at   ZIK   HIKE   (Center   of   Innovation   for   Humoral   Immune Responses in Cardiovascular  Disorders).  Ina  Buchholz  investigates  blood-  and  non-blood  proteins  involved  in autoimmune  diseases  and  the  impact  of  environmental  factors  (e.g.  pH,  ionic  concentrations, temperature),    mutations    and    post-translational    modifications    on    protein    structure    and immunogenicity

Abstract:

Beta2-glycoprotein I (beta2GPI) is a soluble blood protein (326 AA, 5 domains) exhibiting two main conformational states: the circular or closed conformation, where the first domain (DI) is bound  to  the  last  domain  (DV)  of  the  protein;  and  the  linear  or  open  conformation.  In  the  open form,  beta2GPI  binds  to  phospholipid  membranes  via  DV  and  this  form  is  considered  to  play  a crucial role in the autoimmune disease antiphospholipid syndrome (APS). Therefore, investigating the  structural  dynamics   of  this  protein   is  of   high  interest.  We  investigated   different   post- translational  modifications  (PTM)  of  beta2GPI  and  studied  the  impact  on  its  conformation  with biophysical  tools  (e.g.  atomic  force  microscopy,  circular  dichroism  spectroscopy).  Additional insights into the interaction of DI and DV were gained from molecular dynamic simulation studies. PTM  1:  Lysine  residue  acetylation  reveals  a  partial  opening  of  beta2GPI  dependent  on  the acetylation ratio used (Buchholz et al., PCCP 2018). These data indicate that lysines predominantly stabilize the closed conformation and in vivo acetylation via acetyltransferases could destabilize the closed form, leading to a facilitated opening of the structure. PTM 2: Enzymatic reduction of the C- terminal  Cys288/Cys326  disulfide  bond  near  the  putative  contact  interface  of  DI  and  DV  also initiates a conformational change of beta2GPI. Furthermore, disruption of this disulfide bond leads to loosening of a 22 AA flexible loop carrying lysine residues critical for phospholipid membrane binding.  In  summary,  these  PTM  reveal  a  critical  level  of  destabilization of  the  closed  beta2GPI conformation and beta2GPI conformational change may have a large impact on APS disease.

Biography:

Payal Ghosh has pursuing her MS from University of South Florida from College of Pharmacy. She has completed her Bachelor’s degree in Pharmacy from India. Then she worked as a Marketing Manager in a pharmaceutical company. She wants to work on neurological disorder in upcoming future.

Abstract:

alence of neurodegenerative disorders has dominated humankind throughout history. Huntington’s disease (HD) is a prime example in this category. Statistical data shows that every seven to eight people out of 100 are affected by this disease in Western countries. HD causes brain retardation, dementia, and slow movement of limbs and in extreme the death, all of which is primitively caused by damage in brain cells. Though HD is well known, mostly only genetic treatments have been performed till date. Whereas an insufficient production of thyroid hormones can lead to death of brain cells, which results in the occurrence of brain diseases. Dietary iodine acts on thyroid gland and produces the T3 (triiodothyronine) and T4 (thyroxine) to nourish the brain cells, which will prevent the death of brain cell simultaneously the HD. Bilayer lipid coated iodine nanoparticle (outside) along with T3 and T4 (inside) will be administered to the brain. The MCT8 transporter (monocarboxylate 8 transporter) which transports T3 and T4 to the brain will be coated outside of this bi-layer nano-particle. Scanning electron microscopy (SEM) and fluorescence spectroscopy and in vitro analysis on animals will be the best analytical tools. The MCT8 layer will attach to the blood brain barrier (BBB) and makes easy passage for the nano-particle. Nano formulation of iodine acts on brain cell to stimulate its function which will prevent the death of brain cell thus it will slower down the occurrences of the HD. Iodine nanoparticle is a economical and easily available compound and can be used as a good therapeutic agent for HD. As there is no previous data available of the same it will be a great opportunity to investigate this, however, the toxicity level of iodine nanoparticles are still need to be investigated if administered for a long period of time.

Biography:

Ana Merino has completed her PhD at the University of Córdoba (Spain) and Post-doctoral studies at the Biomedical Institute of Bellvitge (IDIBELL) in Barcelona (Spain). She has published more than 30 papers in indexed journals. As a senior Researcher, she is currently developing the project: Pre-clinical evaluation of membranes particles from mesenchymal stem cells for immune and degenerative disorders. This project is being developed in the Internal Medicine department from Erasmus Medical Center (Rotterdam, The Netherlands) in collaboration with the company Takeda-Tigenix.

Abstract:

Mesenchymal stromal cells (MSC) are studied as an immunomodulatory and regenerative therapy in organ repair. Recent work demonstrated that dead MSC which are unable to secret factors, are effective in a sepsis model, suggesting that MSC activity is dependent on the cell membrane interactions with immune cells. We propose a new therapy based on the generation of plasma Membrane Particles (MP) from MSC. We previously showed that MP were effective in reducing the inflammatory phenotype of monocytes. In this study, we investigated the therapeutic potential of MP as regenerative treatment for EC in a TNF-α inflammatory condition. MP were generated from MSC by hypotonic shock and extrusion. MP showed vesicle shape (cryoelectron-microscopy) and a size below 200 nm. Uptake of MP by EC was analyzed by confocal microscopy, and within 24 hours >90% of EC have taken up MP. Three different concentrations of MP were tested on EC. None of the MP concentrations induced apoptosis or activation of EC measured by the expression of the adhesion markers such as ICAM-1, and VCAM. With respect to the regenerative capacity of EC after MP treatment, we have observed an enhancement of angiogenesis by increasing the number of tubes, and branches formation in vitro compared to the negative control (non-treated EC). In the scratch wound healing assays, MP had a stimulating effect on EC to fill the scratch in a dose-dependent manner. In conclusion, MP potentially serve as a novel cell derived therapy that restores vascular integrity and induces endothelial regeneration.

Biography:

Una Janke has studied Humanbiology at the University of Greifswald, with the main focus on Immunology. Currently, she is a PhD student in the group of Prof. Mihaela Delcea at the Institute of Biochemistry and uses biophysical tools to investigate the impact of mutations and environmental factors (e.g. ions, drugs) on activation and immunogenicity of platelet receptor integrin αIIbβ3.

Abstract:

The transmembrane protein integrin alpha IIb beta 3 (αIIbβ3) is involved in hemostasis, wound healing and clot formation. Intracellular as well as extracellular signals can cause inside-out or outside-in signaling, which leads to at least three different conformations: the bent (resting) state; the intermediate extended form; and the ligand-occupied active state. The conformational dynamics of the overall structure of αIIbβ3 during the activation process is possibly related to changes in protein secondary structure, which has not been studied until now in a membrane environment (e.g. liposomes). Moreover, αIIbβ3 is related to the autoimmune disease immune thrombocytpenia, where potential external triggers influence the antigenicity of the integrin by changing the protein structure. In this study we determined the drug-induced activation of αIIbβ3 and the relation to the structure of this protein reconstituted into liposomes. The combination of activation assays and the biophysical tools quartz crystal microbalance, surface plasmon  resonance and circular dichroism spectroscopy show binding of the conformation-specific antibody PAC-1 (which recognizes the active integrin) to αIIbβ3-treated with clinically relevant drugs (e.g. quinine). However, insignificant changes in protein secondary structure were found. Molecular dynamics simulation (MDS) studies confirmed a globular hinge motion in the ectodomain of the integrin with minor changes in protein secondary structure. Our biophysical setup in combination with MDS can be applied to study transmembrane proteins under different conditions in a biomimetic system

Biography:

Perry Habib Saifullah,  I work in the department of Chemistry,  College of science for women,  University of Baghdad.  . The field of my research is based on the diagnosis,  prognosis and,  treatment of diabetes mellitus  patients.  I had supervised  my master and doctorate in the field of biochemistry,  and medical biochemistry  . I also work as a director for Career and development unit in my college and trying to improve the curriculum of our department  in a way that provides students with knowledge skills and soft skills.

Abstract:

Both Alzheimer’s disease (AD) and diabetes mellitus disorder share the most prevalence devastating healthy problems in the old people. Diabetic is a known as a risk factor for the emergence of cognitive dysfunction and dementia complication. Biological and epidemiological evidences confirmed a link between the two diseases, but the precise mechanisms that involved in the developing of cognitive impairment in diabetic patients are not fully comprehended. The Alpha -7 nicotinic acetylcholine receptors (α7-nAChRs) are a type of transmembrane ligand, that are activated through in response to the neurotransmitters such acetylcholine (ACh) and other agonist like nicotine. It belongs to superfamily of receptors that are the fast ionotropic cationic ligand-gate ion channels in nervous, and muscle cell tissues. This receptor play a vital role in the supporting the cognitive function of animal , learning and memory potency by the neurotransmission adjusting , through the control secretion of a different neurotransmitters. Several previous studies had been demonstrated, that the decline in nicotinic receptors mostly the (α-7nAchR) subtype are linked with aging, and also reduced particularly in brain AD patients. The current study included two aims, the first was the investigation about the abundance of (α-7nAchR) at the brain tissue of mice that induced diabetic disease conditions. Second aim was the study the influence of orally metformin drug (which considered as the first line treatment of diabetic disease type 2) on the (α-7nAchRs) abundance in brain tissue of treated diabetic mice by metformin during a therapy duration was 45 days, by means of using the technique detection, the immunohistochemistry (IHC).

Biography:

Eman A Bseiso is working as Assistant Lecturer in the Department of Pharmaceutics & Industrial Pharmacy, Faculty of Pharmacy, October 6 University in Egypt. She earned a Bachelor’s degree in Pharmaceutical Science in 2010 and Master’s degree in Phramaceutics from Faculty of Pharmacy, Ain Shams University in Egypt. Currently, she is pursuing her PhD in Pharmaceutical Technology, Faculty of Pharmacy from the same university. She has published many papers in reputed journals and her current research interests focuses on dermal & intranasal drug delivery systems by using polymers and surfactants to enhance the performance of the large and small drug candidates in addition to nanoparticle formulations for drug targeting (e.g. SLN, NLC & liposomes).

Abstract:

The objective of the current study was to assess the feasibility of nose to brain delivery of melatonin (MEL) using a lipidic nanocarrier (LNCs) for treatment of oxidative diseases. This carrier was chosen owing to its small size and biocompatible nature and is prepared using solvent-free phase inversion temperature technique. Design of experiment was used to establish D-optimal mixture design to study the impact of individual and combined effects of three independent variables; X1 (Oil), X2 (Surfactant), and X3 (Drug), on three LNCs responses; Y1 (particle size), Y2 (Polydispersity index) and Y3 (Zeta potential). MEL-LNCs were successfully prepared and characterized in terms of size, PDI and zeta potential. In-vitro drug release profiles and viscosity were assessed for the optimized LNCs. Results showed that all LNCs had a particle size ranging from 20-200 nm which was appropriate for intranasal permeability and brain targeting. They mostly exhibited narrow size distribution and a negative charge. They also exhibited a sustained release behavior. LNCs were shown to be a promising delivery system for possible nose to brain delivery of MEL.

Biography:

Abstract:

Background: Methotrexate is a systemic immunosuppressant drug that is used for the treatment of psoriasis and psoriatic arthritis. Previous studies demonstrated a potential association between psoriasis and diabetes mellitus, obesity, atherosclerosis and hypertension.

Objective: This study aimed at exploring the possible hyperglycaemic effects of methotrexate in psoriatic arthritis patients (PsA).

Methods: In this prospective cross-sectional study, 27 patients with PsA were evaluated. The status of PsA and presence of accompanying metabolic syndrome was determined by standard criteria and indexes. Blood indicators including, HbA1c, erythrocyte sedimentation rate (ESR), fasting blood sugar (FBS), total cholesterol, high-density lipoprotein (HDL), triglycerides, and C-reactive protein (CRP) were examined before and 12 weeks after methotrexate therapy.

Results: There were no significant changes between HbA1c levels before and after methotrexate therapy in both genders (men: P= 0.131, women: P=0.803). In addition, HbA1c levels in PsA patients with metabolic syndrome were not different before and after treatment (P=0.250). Finally, HbA1c levels did not change in PsA patients without metabolic syndrome before and after therapy (P=0.506).

Conclusion: Methotrexate in PsA patients does not appear to have hyperglycaemic effects in the short-term and can be safely used in patients with metabolic syndrome and diabetes.

Biography:

Nitin Tyagi earned his MBBS degree from Vardhman Mahavir Medical College & Safdarjung Hospital.He is currently the 3rd year post graduate student (MD,Biochemistry)at Department of Biochemistry ,VMMC & SJH,New Delhi.He has authored  7 articles in medical journals.His interests are in cardiology, oncology and  medical genetics

Abstract:

Coronary artery disease(CAD) is one of the most common cardiovascular diseases and is a major cause of morbidity and  mortality worldwide.. Various studies have been done to investigate the role of ADIPOQ gene in the risk of CAD, yet their results have been inconsistent. So, there is a need of genotype analysis of ADIPOQ gene (rs266729) for further evaluation of association between ADIPOQ gene polymorphism and CAD risk.

The aim of the present study was to evaluate the impact of (rs266729) SNP in the promoter region of the ADIPOQ gene on the occurrence of CAD.

MATERIALS AND METHODS: In this case control study, the study group included 50 patients with angiographically proven CAD as case group and 50 apparently healthy age and sex matched adults as control group, for the genotype (C/G) analysis of ADIPOQ gene(rs266729) by PCR-RFLP using Hha I enzyme.

Case Group: CC 20(40%), CG 16(32%) and GG 14(28%); Control Group: CC 29(58%), CG 16(32%) and GG 5(10%). The frequency of allele C in case group was 56% and 74% in control group. The frequency of allele G in case group was 44% and 26% in control group (p=0.0001). There was statistical significance between the two groups (p=0.0001).

CONCLUSION: Adiponectin gene promoter polymorphism (rs266729)  is involved in the pathogenesis of coronary artery disease

KEYWORDS: Adiponectin, Coronary artery disease

Biography:

Henry M. Sobell completed his studies at Brooklyn Technical High School (1948-1952),  Columbia  College  (1952-1956),  and  the  University  of  Virginia  School  of Medicine (1956-1960).  Instead of practicing clinical medicine, he then went to the Massachusetts  Institute  of Technology (MIT) to join Professor Alexander  Rich in the   Department   of   Biology   (1960-1965),   where,   as   a   Helen   Hay   Whitney Postdoctoral Fellow, he learned the technique of single crystal X-ray analysis.  He then joined the Chemistry Department at the University of Rochester, having been subsequently  jointly  appointed  to  both  the  Chemistry  and  Molecular  Biophysics departments  (the  latter  at  the  University  of  Rochester  School  of  Medicine  and Dentistry),  becoming  a  full  tenured  Professor  in  both  departments  (1965-1993). He is now retired and living in the Adirondacks in New York, USA.

Abstract:

Premeltons are examples of emergent structures (i.e., structural solitons) that  arise  spontaneously  in  DNA  due  to  the  presence  of  nonlinear excitations  in  its  structure.     They  are  of  two  kinds:  B-B  (or  A-A) premeltons  form at specific DNA-regions  to nucleate  site-specific DNA melting.    These  are  stationary  and,  being  globally  nontopological, undergo breather motions that allow drugs and dyes to intercalate into DNA.    B-A  (or  A-B)  premeltons,  on  the  other  hand,  are  mobile,  and being  globally  topological,  act  as  phase-boundaries  transforming  B- into  A-  DNA  during  the  structural  phase-transition.     They  are  not expected to undergo breather-motions.  A key feature of both types of premeltons  is  the  presence  of  an  intermediate  structural-form  in  their central  regions  (proposed  as  being  a  transition-state  intermediate  in DNA-melting and in the B- to A- transition), which differs from either A- or B- DNA. Called beta-DNA, this is both metastable and hyperflexible and   contains   an   alternating   sugar-puckering   pattern   along   the polymer-backbone  combined  with  the  partial-unstacking  (in  its  lower energy-forms)  of  every  other  base-pair.    Beta-DNA  is  connected  to either  B-  or  to  A-  DNA  on  either  side  by  boundaries  possessing  a gradation  of  nonlinear  structural-change,  these  being  called  the  kink and the antikink regions.   The presence of premeltons in DNA leads to a  unifying  theory  to  understand  much  of  DNA  physical-chemistry  and molecular-biology.   In  particular,  premeltons  are  predicted  to  define the  5’  and  3’  ends  of  genes  in  naked-DNA  and  DNA  in  active- chromatin, this having important implications for understanding physical aspects  of  the  initiation,  elongation  and  termination  of  RNA-synthesis during transcription.   For these and other reasons, the model will be of broader interest to the general audience  working in these areas.   The model explains a wide variety of data, and carries within it a number of   experimental   predictions   –   all   readily   testable   –   as   will   be described in my talk.

Biography:

Nicolas studied at the University of Compiegne (France) and completed his Engineer in Biotechnology degree. For his master and PhD in Neuroscience from the University Pierre et Marie Curie, he moved to the Pasteur Institute in Paris, where he worked in the group of Dr. Pierre-Jean Corringer and Prof. Jean-Pierre Changeux (Channel receptors group) on the elucidation of the crystal structure of a pentameric ligand gated ion channel in an open conformation. From 2009 to 2013, Nicolas moved to FMI (Friedrich Miescher Institute for Biomedical research) as a post-doctoral fellow in the group of Dr. Nicolas Thomae, where he worked on the mechanisms of Holliday junction dissolution by solving the structure of the human Topoisomerase III in complex with a modulatory protein called RMI1. From 2013 to 2017, he worked at Roche, first as a Roche post doctoral fellow and after as a scientist in the Chemical biology department developing biophysical methods for membrane proteins as well as producing, purifying, stabilizing and characterizing GPCRs, transporters and membrane enzymes.

Abstract:

Today, soluble proteins are managed routinely within the project timelines and scope with the rapid portfolio changes in pharmaceutical industry. Establishment of biophysical and structure-based methods for transmembrane proteins still represents a significant challenge to have an impact on drug discovery. leadXpro combines membrane protein expression, purification and structure determination coupled to premium access to the synchrotron Swiss Light Source (SLS), the Free Electron Laser (SwissFEL) and single particle cryo-electron microscopy (cryo-EM) at the University of Basel. LeadXpro also confronts  structural data to different biophysical measurements like thermal shift assays, radiobinding assay and wave guide interferometry in order to generate better lead molecules with appropriate features.

The talk/poster will show advancements in projects and technologies with examples for serial crystallography performed at synchrotron and free electron laser enabling structure determination of challenging drug targets. Moreover, recent efforts and implementation of wave-guide interferometry method for analysis of small (fragment-like molecules)/large ligand binding kinetics on membrane proteins will be discussed in the context of i) lead discovery and optimization ii) biologics targeting membrane proteins. Finally, recent progress in cryo-EM will also be discussed.

Biography:

Balasubramani G L has his expertise in computational, molecular biology and structural biochemistry. Holding 7-years of research experience in DNA-binding proteins from Mycobacterium tuberculosis. Also, have profound knowledge in programming languages, computational methodologies and Sun certified JAVA programmer. Through drug-repurposing approaches, he found out drugs which are active against Mycobacterium tuberculosis targets. This approach is novel and inexpensive to deal with drug-resistant strains.

Abstract:

Drug repurposing has gained momentum globally and become an alternative avenue for drug discovery. Though tuberculosis (TB) can be cured with the use of currently available anti-tubercular drugs, emergence of drug resistant strains of Mycobacterium tuberculosis H37Rv (Mtb) and the huge death toll globally, together necessitate urgently newer and effective drugs for TB. To address this problem, we screened FDA-approved drugs by virtual screening and binding free energy calculations to identify novel inhibitors that target two active sites of Mtb DNA gyrase (i) dimer interface of gyraseA subunit, (ii) a active site region of ATP binding (N-terminal domain) pocket on gyrase B subunit overlapping the site targeted by coumarin drugs. Here, we identified total of 4 compounds tightly binds to ATPase binding pocket of N-terminal domain of gyrase B. Results obtained from biochemical and biophysical studies shows strong binding of screened compounds and inhibits gyrase catalytic cycle. Our evidence strongly suggests that these compounds bind to the N-terminal domain of gyrase B. Furthermore, we performed drug susceptibility test using screened compounds on Mtb and showed less MICs as compared to reported drugs. This finding indicates all the identified compounds represents potential scaffolds for further optimization of novel antibacterial agents that can act on drug-resistant strains.

Biography:

Oznur Aglar completed her Master studies in Organic Chemistry at Umea University, Sweden, in 2016. She is a second-year PhD doctoral student in Analytical Chemistry at the University of Potsdam. Her current research focuses on the study of protein-ligand  interactions, including binding affinity and binding kinetics by NMR.

Abstract:

Ceramide, an important component in the metabolism of sphingolipids, plays a significant role in proliferation, and apoptosis of cells.1 De novo synthesis of ceramide takes place at the cytosolic surface of the endoplasmic reticulum (ER), and then, ceramide is transferred to the Golgi apparatus for conversion into sphingomyelin and glucosphingolipids, mainly by non-vesicular trafficking. Non-vesicular transport of ceramide is carried out by the ceramide transfer protein (CERT) that consists of peptidic motifs and multiple domains.1 The C-terminal (StAR)-related lipid transfer (START) domain is the most important domain given the fact that it is capable of extracting and accommodating ceramide in its deep hydrophobic cavity. CERT  could be an attractive pharmacological target because of its involvement in common pathological processes such as Alzheimer's disease, infectious diseases and cancer.2 A well-known antagonist of CERT is  -N-(3-hydroxy-1-hydroxymethyl-3-phenylpropyl) dodecanamide (HPA-12),3 however, there is only limited structure-activity relationship (SAR) data available. In this study, we aim to explore the interaction between CERT and HPA-12 to establish SAR of this compound class by nuclear magnetic resonance spectroscopy (NMR) in order to improve the inhibition activity of the ligand for a potential drug design.

Herein, we optimized the established expression and purification protocols to get a good yield of the monomer of the START domain. In order to get an idea about the suitability of the protein of interest for ligand and receptor-based NMR experiments, the target protein was labeled with 15N and purified using an optimized three step purification  method. The 15N labeled START domain’s monomer was used to record initial 1H - 15N HSQC and TROSY NMR spectra in phosphate buffer. Results of both HSQC and TROSY experiments were promising for further receptor based NMR experiments.

Biography:

Ponizovskiy M.R.  Is head of Laboratory Biochemistry and Toxicology at Kiev regional p/n hospital.He has done many of his projects in Biochemistry and Toxicology field, and he has dedicated one of his articles entitled “Biophysical and Biochemical Mechanisms of Forming and Development A Human Eukaryotic Organism from Single Pluripotent Cell into Multicellular Embryo and A Living Organism in Norm in memory of his daughter T.M.Ponizovska..

Abstract:

The mechanism forming and development a human eukaryotic organism from single pluripotent cell was described from the point of view of thermodynamics, biophysics and biochemistry. The metabolic mechanisms of an organism during its life are subjected as to outer influences from surroundings as well as to inner influences. An organism expends some energy from Basic Internal Energy which is stored energy in Basic stem cells (neurons of Central Nervous System) which share energy via sequentially Basic stem cells → Totipotent stem cells → Pluripotent stem cells → Multipotent stem cells → Oligopotent stem cells and then distrib¬uting between cells various types leading to cells’ proliferations. However this expenditure Basic Internal Energy results in senescence of an organism. Just mechanisms of genomic processes activity are the links of mechanisms stem cells operations which cause advance an organism during its life determining duration life of an organism. In a developing embryo, stem cells generate differentiation into all the specialized cells forming corresponding tissues (ectoderm, endoderm, mesoderm etc.).

These transformations of stem cells maintain stability each tissue of an organism, such as blood, skin, intestinal tissues etc. These transformations of stem cells are the potency of obtained energy which specifies differentiation into different cell types of the stem cell. Also there were described mechanisms Gametogenesis, impregnation ovum by sperms and foetus growth from single cell considering flows energy from stem cells to cells types which cause transition diploid cellular division through mitosis into haploid cellular division through meiosis and back into diploid cellular division through mitosis which are subjected to Boltzmann theory of transition microstates into macrostate as increased Entropy into decreased Entropy for saving energy. The genesis and development of an organism was explained using famous Prigogine theorem and Glansdorff and Prigogine theory sharing into such stages of human life development: born of an organism, babyhood, childhood, young age, juvenile age, middle age, full age, elderly age, old age. There was estimated levels metabolic activity of each stage of human life. Also all stages of human life development were considered via energy flow which gener¬ates cells developments through stem cells into cells types. The mechanisms of these transformations cells were de¬scribed from point of views of thermodynamics, biophysics and biochemistry. In a development babyhood and childhood, stem cells exert expression metabolic processes operations stimulating hormonal processes and immune defensive processes.

In a development young age years and juvenile age years, stem cells continue exertion metabolic processes operations stimulating hormonal processes and immune defensive processes as well as forming sex organs, sex characters and Gametogenesis with possible generating foetus cells which receive energy from mother’s stem cells. In a development middle age years and full age years, stem cells continue maintenance stability Internal Energy an organism, its hormonal and immune functions. In elderly age years, stem cells have less energy than in middle age and full age for continue maintenance stability Internal Energy an organism, its hormonal and immune functions. In aging organisms, stem cells have insufficient energy for continue maintenance stability Internal Energy an organism, its hormonal and immune functions.

Biography:

Prakash Kinthada is a Professor in Chemistry at Sri Vidyanikethan Engineering college, JNTU University in Ananthapur, A. Rangam Peta, Tirupathi, India.He has done his Ph.D from Sri Krishna Devaraya University, Anantapur, India during the period  1981-1987.

Abstract:

Cancer is a dreadful disease and any practical solution in combating this disease is of paramount importance to public health. Cancer patients have burdened by drug induced toxic side effects, and no turned to seek help from the complementary and alternative medicine hoping for a better cure. Research on Platinum based drugs and Non Platinum based drugs is a Multi-Million Dollar Industry in USA and there is every need to produce safe drugs for the cure of this monstrous disease. Flavonoids have a long history of use in traditional medicines in many cultures. The phytochemical, curcumin is one of the major dietary flavonoid, belonging to a group of flavonol, Curcumin is a natural polyphenol. It is highly potential molecule capable of preventing and treating various cancers.  Various dietary chemo preventive agents, turmeric powder or its extract are broadly used as therapeutic preparations in Indian System of medicine. We provide a summarized synthesis and structural determination of Curcumin Oxime, Curcumin Thiosemicarbazone derivative of Gold (III) complex. The use of these analogs for prevention of cancer tumor progression and treatments of human malignancies. A pharmacologic agent for treating and/or preventing cancer, among other diseases and conditions, and particularly breast, prostate, and pancreatic cancer, in humans and animals. The novel pharmacologic agent is an isoflavonoid or isoflavonoid mimetic covalently attached to a cytotoxic pharmacophore that, preferably has the ability to conjugate with a metal salt to form a more potent metal complex, particularly a Au (III) complex and other complexes of Platinum, Palladium, Ruthenium, Copper etc.

Biography:

Amir Zeb is PhD student at Geyongsang National University, South Korea. His research interest is Computer Aided Drug Designing and Molecular Modeling. Mr. Zeb has been exposed to a number of proteins modeling projects and achieved excellent output. Currently, Mr. Zeb is trying to unveil the mechanistic studies of therapeutics targets in neurological disorders and their computational inhibition. He has published more than 15 peer reviewed articles in reputed journals.

Abstract:

Neurotoxic insults activate calpain, which in turn produces truncated p25 from p35. p25 forms hyperactivated Cdk5/p25 complex, and thereby induces severe neuropathological aberrations including hyperphosphorylated tau, neuroinflammation, apoptosis, and neuronal death. Inhibition of Cdk5/p25 complex alleviates aberrant phosphorylation of tau to mitigate AD pathology. PHA-793887 and Roscovitine have been investigated as selective inhibitors of Cdk5/p25 with IC50 values 5nM and 160nM, respectively, but their mechanistic studies remain unknown. Herein, computational simulations have explored the binding mode and interaction mechanism of PHA-793887 and Roscovitine with Cdk5/p25. Docking results suggested that PHA-793887 and Rsocovitine have occupied the ATP-binding site of Cdk5 and obtained highest docking (GOLD) score of 66.54 and 84.03, respectively. Furthermore, molecular dynamics (MD) simulation demonstrated that PHA-793887 and Roscovitine established stable RMSD of 1.09 Å and 1.48 Å with Cdk5/p25, respectively. Profiling of polar interactions suggested that each inhibitor formed hydrogen bonds (H-bond) with catalytic residues of Cdk5 and could remain stable throughout the molecular dynamics simulation. Additionally, binding free energy calculation by molecular mechanics/Poisson–Boltzmann surface area (MM/PBSA) suggested that PHA-793887 and Roscovitine had lowest binding free energies of -150.05 kJ/mol and -113.14 kJ/mol, respectively with Cdk5/p25. Free energy decomposition demonstrated that polar energy by H-bond between the Glu81 of Cdk5 and PHA-793887 is the essential factor to make PHA-793887 highly selective towards Cdk5/p25. Overall, this study provided substantial evidences to explore mechanistic interactions of the selective inhibitors of Cdk5/p25 and could be used as fundamental considerations in the development of structure-based selective inhibitors of Cdk5/p25.

Biography:

Evangelia E. Chronpoulou graduated from the Department of Agricultural Biotechnology (Agricultural University of Athens, Greece) in 2005, she received her Master degree from the Medical School of the University of Athens in 2008 and her PhD in Enzyme Biotechnology from Agricultural University of Athens in 2013. Currently, she is carrying out postdoctoral research at the Agricultural University of Athens. Evangelia has co-authored more than 25 peer-reviewed articles in peer-reviewed journals and 9 chapters in scientific books.

Abstract:

In the last few decades there has been a considerable increase in demand for enzymes as biocatalysts in research, pharmaceutical, food and laundry industry, biofuel development, agroindustry and fishery waste exploitation.1 Protein engineering is a prominent method to overcome limitations that can be faced in these areas1, aiming to the design and creation of new enzymes with novel of improved catalytic and structural features.2 Enzyme nanostructures are considered as an effective materials for the development of assembled structures with complexity and functional diversity.3,4  GSTs are multi-functional enzymes of high-importance for a number of biotechnological applications. They are considered as good model systems for engineering studies, due to their modular catalytic and binding features.5 This work was inspired by natural protein nanostructures and “lego’’ chemistry, aiming to the  design and development of novel protein folds that can be self-assembled into complex-structures by exploiting the modularity capabilities of human GSTA1-1 enzyme (hGSTA1-1).Based on the analysis of the crystal structure of hGSTA1-1 (PDB identification code: 1K3Y)6, the residues Lys140 and Ser141were selected as potential hot-spots for step-by-step site-saturation mutagenesis. Four novel enzymes were designed and constructed: hGSTA1-1G (K140H), hGSTA1-1Ε (K140H, S141H), hGSTA1-1S (with the motif EKVLH before the mutation Κ140Η) and  hGSTA1-1B (with more repetitive motives EKVLH after PCR-ligation). These GST genes were cloned in 5EXP-CT TOPO vector and expressed in E. coli. The recombinant enzymes were purified by affinity chromatography and their biochemical properties were studied. The results showed that the mutant and “lego” enzymes appear to exhibit improved catalytic efficiency and selectivity towards pesticides, without loosing their structural stability. The discovery and characterization of such enzymes, with the ability to fold and retain catalytic activity, can provide future building blocks in protein engineering efforts.

Biography:

James Endrizzi, a Ph.D. chemist, specializes in the use X-ray crystallography to unveil  macromolecular function.  With a B.S. in Chemistry & Biology from the U. of Minnesota and a doctorate from  the U. of Oregon, James went on to post-doctoral work at UC Berkeley.  After 9 years at Berkeley, he went on to research stints at UC Davis, Montana State University and the Hormel Institute.  Along the way, Dr. Endrizzi has presented 17 talks and over 20 posters at conferences while  publishing 24 papers in top tier jornals.  In 2012, he founded SuperNova C, a company devoted to providing vitamin C as mineral ascorbates.

Abstract:

Aspartate  transcarbamoylase (ATCase),  a textbook case for allosteric enzyme  regulation, catalyzes the first step of pyrimidine nucleotide biosynthesis and is feedback inhibited by pathway end-product cytidine triphosphate (CTP). ATCase exhibits cooperative substrate binding to its’ catalytic subunits and allosteric binding of nucleotides to its’ regulatory subunits.  Inhibition by CTP and activation by ATP  promote homeostasis of purine and pyrimidine nucleotides.  Regulation of ATCase activity is interpreted as a ligand-promoted change in the equilibrium between inactive (taut) and active (relaxed) states, consistent with a two-state allosteric model.  In this view, ligand-free enzyme represents the inactive, taut state while the structure of ATCase bound to bisubstrate analog N-phosponacetyl-L-aspartate (PALA) represents the active, relaxed  state. However, there is little evidence that PALA-bound ATCase represents the active conformation(s) in the absence of substrates.  A novel approach to define the structure of the ligand-free relaxed, or “activated” state is to use enzyme variants that destabilize the taut state and shift the allosteric equilibrium toward the relaxed-state conformation(s). In contrast to the wild-type enzyme, which exhibits a more compact global conformation and sigmoidal enzyme kinetics, the mutants are more expanded and display hyperbolic kinetics characteristic of non-cooperative enzymes.  We tested the hypothesis that ATCase is activated through modulation of flexibility by determining the X-ray crystal structures of activated-state  mutants. Multiple crystal forms were identified for 3 variants.  We have refined dozens of structures thus far  Myriad tertiary and quaternary structures observed for “active” variants supports the hypothesis that these structures represent metastable states accessed by an ensemble of activated-state conformations.  

Biography:

Mahesh Chandra Patra is currently pursuing PhD degree at Ajou University, Suwon, Korea in the field of computational biology. He holds a Master’s degree in Bioinformatics from Orissa University of Agriculture and Technology, Bhubaneswar, India. He has expert level knowledge of molecular dynamics simulations and python programming language. He has published 25 papers including two review articles in reputed journals. Currently, he is developing a database for therapeutically relevant clinical/preclinical agents of innate 

Abstract:

The Toll/interleukin 1 receptor (TIR) domain-containing adaptor protein (TIRAP) regulates Toll-like receptor (TLR) 2, TLR4, TLR7, and TLR9 signaling pathways. TIRAP anchors to phosphatidylinositol (PI) 4,5-bisphosphate (PIP2) on the plasma membrane and PI (3,4,5)-trisphosphate (PIP3) on the endosomal membrane and assists in recruitment of the myeloid differentiation primary response 88 protein to activated TLRs. To date, the structure and mechanism of TIRAP's membrane association are only partially understood. Here, we modeled an all-residue TIRAP dimer using homology modeling, threading, and protein-protein docking strategies. Molecular dynamics simulations revealed that PIP2 creates a stable microdomain in a dipalmitoylphosphatidylcholine bilayer, providing TIRAP with its physiologically relevant orientation. Computed binding free energy values suggest that the affinity of PI-binding domain (PBD) for PIP2 is stronger than that of TIRAP as a whole for PIP2 and that the short PI-binding motif (PBM) contributes to the affinity between PBD and PIP2. Four PIP2 molecules can be accommodated by distinct lysine-rich surfaces on the dimeric PBM. Along with the known PI-binding residues (K15, K16, K31, and K32), additional positively charged residues (K34, K35, and R36) showed strong affinity toward PIP2. Lysine-to-alanine mutations at the PI-binding residues abolished TIRAP's affinity for PIP2; however, K34, K35, and R36 consistently interacted with PIP2 headgroups through hydrogen bond (H-bond) and electrostatic interactions. TIRAP exhibited a similar interaction pattern and binding affinity with PIP3 as it did with PIP2 through an H-bond network involving K34, K35, and R36. The present study enabled us to understand the mechanism of TIRAP's membrane association that can be useful for designing peptide-based drugs targeting TLR2-, TLR4-, TLR7-, and TLR9-mediated autoimmune diseases

Biography:

Sergio Senar has completed his PhD at the age of 28 years from Alcalá de Henares University and postdoctoral studies on GPCR orphan receptors from SmithKline Beecham Natural Products Research Department. After a long career supporting early drug discovery at GSK he became self-employed at DrTarget, a non-profit Bio-Soft service entity devoted to consultancy on early portfolios for academy and pharma industry. He has published articles on signal transduction, drug discovery and mechanistic biochemistry.

Abstract:

The use of computational tools in the early stages of drug development has increased in recent decades. Machine learning approaches have been of special interest , since they can be applied in several steps of the drug discovery methodology, such as prediction of target structure, prediction of biological activity of new ligands through model construction, discovery or optimization of hits, construction of models that predict the pharmacokinetic and toxicological (ADMET) profile of compounds, and assessment of mechanism of action and identification of new targets for further research. .

Two public domains, NCBI and ChemBl hold millions of experimental records accessible to the whole biomedical community. The websites support applications to facilitate access to results for limited number of assays or molecules, but they also allow downloads of the database components, sometimes, of the database itself.

In this conference we will  present an overview on some applications of ML techniques in construction of classification and/or prediction models of biological activity, identification of mechanism of action of molecules active in phenotypic assays and unveiling of potential new targets through pathways analysis. By carrying out a virtual screening on 1,.5M compounds, we´ll apply the results to the repurposing of marketed drugs to new therapies and match predicted drug activity to gene expression or occurrence of mutations. This last approach being of particular interest for the identification of combination of therapies in cancer

Biography:

Dr.Abdul Rouf Mir is working as a Senior Resident in the Department of Biochemistry, Faculty of Medicine, Aligarh Muslim University, Aligarh, India. He has completed PhD program from the same department and has published 20 research papers/reviews in peer reviewed international journals. He has presented his work at various national and international conferences, and is a reviewer to international journals. He has qualified the eligibility test for Assistant Professorship and was awarded MANF-JRF and MANF-SRF by University Grants Commission. He has taught graduate students at the College affiliated to University of Kashmir.

Abstract:

Increased oxidative stress under hyperglycemic conditions, through the interaction of AGEs with RAGE receptors and via activation of interleukin mediated transcription signalling, has been reported in cancer. Proteins modifications are being explored for their roles in the development and progression of cancer and autoantibody response against them is gaining interest as a probe for early detection of the disease. This study has analysed the changes in histone H1 upon modification by methylglyoxal (MG) and its implications in auto-immunopathogenesis of cancer. Modified histone showed modifications in the aromatic residues, changed tyrosine microenvironment, intermolecular cross linking and generation of AGEs.  It showed masking of hydrophobic patches and a hypsochromic shift in the in ANS specific fluorescence. MG aggressively oxidized histone H1 leading to the accumulation of reactive carbonyls. Far UV CD measurements showed di-carbonyl induced enhancement of the alpha structure and the induction of beta sheet conformation; and thermal denaturation (Tm) studies confirmed the thermal stability of the modified histone. FTIR analysis showed amide I band shift, generation of a carboxyethyl group and N-Cα vibrations in the modified histone. LCMS analysis confirmed the formation of Nε-(carboxyethyl)lysine and electron microscopic studies revealed the amorphous aggregate formation. The modified histone showed altered cooperative binding with DNA. Modified H1 induced high titre antibodies in rabbits and the IgG isolated form sera of rabbits immunized with modified H1 exhibited specific binding with its immunogen in Western Blot analysis. IgG isolated from the sera of patients with lung cancer, prostate cancer, breast cancer and cancer of head and neck region  showed better recognition for neo-epitopes on the modified histone, reflecting the presence of circulating autoantibodies in cancer. Since, reports suggest a link between AGE-RAGE axis and carcinogenesis, glycoxidation of histone H1 and its immunogenicity paves ways for understanding role of glycoxidatively damaged nuclear proteins in cancer

Biography:

Shuntaro Hara received his M.S. and Ph.D. degrees from the University of Tokyo and started his research career as a postdoctoral fellow at the University of Tokyo. After postdoctoral training, Dr. Hara has worked as a research associate at the National Cardiovascular Center Research Institute (Osaka, Japan), as a visiting fellow at University College London, and as an assistant professor at Kitasato University (Tokyo, Japan), and then became a professor at the School of Pharmacy, Showa University in 2009. He has published more than 100 papers in reputed journals.

Abstract:

Acyl-coenzyme A synthetase long-chain family members (ACSLs) are a family of enzymes that convert long-chain free fatty acids into their acyl-CoAs. Among ACSL isozymes, ACSL4 has been hypothesized to modulate the metabolic fates of polyunsaturated fatty acids including arachidonic acid. In the present study, to investigate the enzymatic and protein characteristics of ACSL4, the cDNA for human ACSL4 was cloned from human epithelial colorectal adenocarcinoma Caco-2 cells and then recombinant ACSL4 enzyme containing a C-terminal His-tag was expressed in Spodoptera frugiperda 9 (Sf9) cells using the baculovirus expression system. ACSL4 enzyme activity was detected in 10,000 x g supernatants of ACSL4-expressing Sf9 cell lysates and then partially purified by nickel affinity column chromatography. We further investigated the substrate specificity of recombinant human ACSL4 by LC/MS and found that ACSL4 enzyme preferred various kinds of polyunsaturated fatty acid including docosahexaenoic acid, docosapentanoic acid, eisopentaenoic acid, and dihomo-γ-linolenic acid, as well as arachidonic acid as a substrate. On the other hand, oleic acid, linoleic acid and linolenic acid were poor substrates, although these fatty acids contain unsaturated bonds. These results confirmed the importance of ACSL4 in maintenance of membrane phospholipid bearing polyunsaturated fatty acid.

Biography:

Shuntaro Hara received his M.S. and Ph.D. degrees from the University of Tokyo and started his research career as a postdoctoral fellow at the University of Tokyo. After postdoctoral training, Dr. Hara has worked as a research associate at the National Cardiovascular Center Research Institute (Osaka, Japan), as a visiting fellow at University College London, and as an assistant professor at Kitasato University (Tokyo, Japan), and then became a professor at the School of Pharmacy, Showa University in 2009. He has published more than 100 papers in reputed journals.

Abstract:

Acyl-coenzyme A synthetase long-chain family members (ACSLs) are a family of enzymes that convert long-chain free fatty acids into their acyl-CoAs. Among ACSL isozymes, ACSL4 has been hypothesized to modulate the metabolic fates of polyunsaturated fatty acids including arachidonic acid. In the present study, to investigate the enzymatic and protein characteristics of ACSL4, the cDNA for human ACSL4 was cloned from human epithelial colorectal adenocarcinoma Caco-2 cells and then recombinant ACSL4 enzyme containing a C-terminal His-tag was expressed in Spodoptera frugiperda 9 (Sf9) cells using the baculovirus expression system. ACSL4 enzyme activity was detected in 10,000 x g supernatants of ACSL4-expressing Sf9 cell lysates and then partially purified by nickel affinity column chromatography. We further investigated the substrate specificity of recombinant human ACSL4 by LC/MS and found that ACSL4 enzyme preferred various kinds of polyunsaturated fatty acid including docosahexaenoic acid, docosapentanoic acid, eisopentaenoic acid, and dihomo-γ-linolenic acid, as well as arachidonic acid as a substrate. On the other hand, oleic acid, linoleic acid and linolenic acid were poor substrates, although these fatty acids contain unsaturated bonds. These results confirmed the importance of ACSL4 in maintenance of membrane phospholipid bearing polyunsaturated fatty acid.