Keynote address by Prof. Amitabha Chattopadhyay

Prof. Amitabha Chattopadhyay received B.Sc. with Honors in Chemistry from St. Xavier’s College (Calcutta) and M.Sc. from IIT Kanpur. He obtained his Ph.D. from the State University of New York (SUNY) at Stony Brook, and was a Postdoctoral Fellow at the University of California, Davis. He subsequently joined the Centre for Cellular and Molecular Biology (CCMB) in Hyderabad and now is an Outstanding Scientist (Director Level) there.

Prof. Chattopadhyay’s work is focused on monitoring organization, dynamics and function of biological membranes in healthy and diseased conditions. His group has developed and applied novel, innovative and sensitive techniques (such as the wavelength-selective fluorescence approach) using fluorescence spectroscopy for monitoring solvent relaxation in membranes, membrane-mimetic media, and proteins. These pioneering studies have led to a better understanding of the dynamics of hydration of membranes and proteins. Another seminal contribution of Prof. Chattopadhyay’s group focuses on the role of membrane cholesterol in regulating the organization, dynamics and function of G protein-coupled receptors such as the serotonin1A receptor. His work showed, for the first time, that membrane cholesterol is necessary for the function of G protein-coupled receptors such as the serotonin1A receptor. His work has also provided novel insight in the role of membrane cholesterol in the entry of pathogens into host cells. Prof. Chattopadhyay has used fluorescence-based microscopic approaches such as Fluorescence Recovery After Photobleaching (FRAP), Fluorescence Correlation Spectroscopy (FCS), and Fluorescence Resonance Energy Transfer (FRET) to provide novel insight into organization, dynamics and function of membrane-bound receptors. Overall, his work has contributed significantly to the understanding of membrane organization and dynamics, and the interplay between membrane lipids and proteins, especially in neuronal membranes.

Prof. Chattopadhyay was awarded the prestigious Shanti Swarup Bhatnagar Award, Ranbaxy Research Award, Prof. G.N. Ramachandran 60th Birthday Medal from the Indian National Science academy, and is a J.C. Bose Fellow of the Dept. of Science and Technology, Govt. of India. He is an elected Fellow of the Royal Society of Chemistry, and all the Indian Academies of Science, the Telangana Academy of Sciences, and West Bengal Academy of Science and Technology. Prof. Chattopadhyay has served on the editorial boards of a large number of reputed international journals that include Biophysical Journal, The Journal of Physical Chemistry, Journal of Neurochemistry, BBA-Biomembranes, Journal of Membrane Biology, FEBS Letters, IUBMB Life and ACS Chemical Neuroscience. He has mentored a number of students for Ph.D. Prof. Chattopadhyay has authored more than 200 research publications (mostly as first or senior/corresponding author; total citations > 8500, h-index 48, i-10 index 161), a monograph, and national and international patents. He has delivered more than 500 invited lectures all over the world including keynote, plenary, and colloquium lectures. Prof. Chattopadhyay has organized a number of international conferences on the broad theme of biological membranes including a thematic meeting of the Biophysical Society. Prof. Chattopadhyay has been instrumental in designing and teaching courses related to biomembranes and fluorescence spectroscopy for Ph.D. students in India and other parts of the world. In recent years, Prof. Chattopadhyay has been involved with science awareness programs among high school and college students. Prof. Chattopadhyay is an Adjunct Professor at the Tata Institute of Fundamental Research (Mumbai), Indian Institute of Technology (Kanpur), Jawaharlal Nehru University (New Delhi), Indian Institute of Science Education and Research (Mohali), Royal Melbourne Institute of Technology (Australia), Swinburne University of Technology (Australia), and Honorary Faculty at the Jawaharlal Nehru Centre for Advanced Scientific Research (Bangalore). He serves as the first Dean of Biological Sciences of the Academy of Scientific and Innovative Research. Session – I Talk: 1

Name of the Speaker: Dr. Anand Ballal, SO/G, Molecular Biology Division, Bhabha Atomic Research Centre (BARC), Mumbai.

Time: 10:00 – 10:30am

Life Amidst Oxidative Stress: Role of Catalases in Photosynthetic Bacteria Anand Ballal1*, Dhiman Chakravarty1, Manisha Banerjee1 and Subhash C Bihani2

The photosynthetic, filamentous, N2-fixing bacterium, Anabaena, is one of the few species in the world that uses solar energy (i.e. photosynthesis) to convert the inert atmospheric N2 into bio- + utilizable NH4 . For this reason, Anabaena is widely used as an eco-friendly biofertilizer in the paddy fields of Asia. Photosynthesis and nitrogen-fixation are both sensitive to oxidative stress, which is mediated by the toxic reactive oxygen species (ROS). Our research has focused on the role played by the anti-oxidant enzymes, catalases and peroxidases, in overcoming oxidative stress in Anabaena. Recent advances from our laboratory that highlight (1) the physiological role of catalase under conditions of H2O2/salinity stress and (2) the novel molecular structure of catalase will be discussed.

Talk: 2

Name of the Speaker: Dr. Sanjeev Srivastava, Associate Professor, Department of Biosciences and Bioengineering, Indian Institute of Technology (IIT), Bombay.

Time: 10:30 – 11:00am

Basics of Proteomics: A case study on Malaria for Identification of Potential Prognostic Serum Biomarkers and understanding pathobiology of disease

Plasmodium vivax malaria is no longer benign due to the parasite’s ability to elicit severe symptoms even at very low parasitic biomass and the frequent occurrence of relapses, months after the clearance of primary infection. However, the mechanisms underlying the pathogenesis of vivax malaria still remain obscure with limited knowledge on its biology. In our study, serum samples from patients across different endemic regions of India were comprehensively investigated using a multiomics approach including quantitative proteomics platforms like DIGE and iTRAQ and a metabolomics approach using LCMS to identify markers of disease severity. Functional pathway analysis of the differentially expressed proteins revealed modulation of several vital physiological pathways, including anti-oxidative stress pathways, lipid metabolism, complement cascades and blood coagulation in vivax malaria. Proteins such as Carbonic anhydrase, Superoxide dismutase 1, Fatty acid-binding protein, nebulin, profilin, Serum amyloid A, Haptoglobin, Apo A-I and Apo E exhibited sequential alterations in their expression levels in different severity levels of malaria. The results were validated using ELISA and SPR-based measurements. An MRM-based methodology using LCMS-8050 was optimized for proteins such as Ceruloplasmin, Alpha 1-acid glycoprotein and Alpha 1-antichymotrypsin using samples from both falciparum and vivax malaria patients. This approach will be further explored for the validation of other interesting targets of vivax malaria. Furthermore, the first ever comparative metabolomic analysis of severe and non-severe vivax malaria was performed to gain a deeper insight into disease pathogenesis. Using LC-MS coupled with multivariate statistical data analysis approaches over 3000 serum metabolites were screened. Metabolites involved in oxidative stress such as nitrotyrosine, tyrosine etc. were found to be upregulated in vivax malaria patients in accordance with the proteomics data for oxidative protein markers which were further validated using kit-based assays. We conclude that the multiple serum proteins, antioxidative enzymes and oxidation protein products cumulatively represent the oxidative stress and antioxidative status of the patients with malaria and reflecting the level of disease severity.

Talk: 3

Name of the Speaker: Dr. Raghavendra Patvardhan, Scientific Officer D, Free Radical Biology Section, Radiation Biology & Health Science Division, Bhabha Atomic Research Centre (BARC), Mumbai.

Time: 11:00 – 11:20am

Mitigation of radiation induced hematopoietic injury via regulation of cellular MAPK/phosphatase levels and increasing hematopoietic stem cells

Here we describe a novel strategy for mitigation of ionizing radiation (IR) induced hematopoietic syndrome by suppressing the activity of MKP3 resulting in ERK activation and enhanced abundance of hematopoietic stem cells using an antioxidant flavonoid baicalein (5,6,7trihydroxyflavone). It offered complete protection to mouse splenic lymphocytes against radiation induced cell death. Inhibitors of ERK and Nrf-2 could significantly abrogate baicalein mediated radioprotection in lymphocytes. Baicalein inhibited phosphatase MKP3 and thereby enhanced phosphorylation of ERK and its downstream proteins like Elk and Nrf-2. It also increased the nuclear levels of Nrf-2 and mRNA levels of its dependent genes. Importantly, baicalein administration to mice prior to radiation exposure led to significant recovery in loss of bone marrow cellularity and also inhibited cell death. Administration of baicalein increased the hematopoietic stem cell frequency as measured by side population assay and also by antibody staining. Further, baicalein offered significant protection against whole body irradiation (7.5 Gy) induced mortality in mice. Interestingly, we found that baicalein works by activating same target molecules ERK and Nrf-2 both in vitro and in vivo. Finally, administration of all-trans-retinoic acid (inhibitor of Nrf-2) significantly abrogated baicalein mediated protection against WBI induced mortality in mice. Thus, in contrary to the generalized conception of antioxidants acting as radioprotectors, we provide a rationale that antioxidants exhibit pleiotropic effect through activation of multiple cellular signaling pathways.

Session – II Talk: 1

Name of the Speaker: Dr. Roop Malik, Associate Professor and Wellcome – DBT Senior Fellow, Department of Biological Sciences, Tata Institute of Fundamental Research (TIFR), Mumbai

Time: 11:45am – 12:15pm

Lipolysis in the liver and active transport of lipid: A new connection?

The liver is a major regulator of lipid flux in the body. How lipid is stored and utilized in the liver in metabolically regulated manner is central to understanding lipid homeostasis and the consequences of its failure such as fatty liver diseases, obesity, diabetes. New evidence from our work shows that the metabolic state of liver is correlated with an unusually high motility of lipid particles along microtubules. The molecular mechanism behind this observation and its implications will be presented. Talk: 2

Name of the Speaker: Dr. Sameer Maji, Associate Professor, Department of Biosciences and Bioengineering, Indian Institute of Technology (IIT), Bombay.

Time: 12:15 – 12:45pm

Amyloids as functional Bio-materials

Amyloids are protein/peptide aggregates mainly associated with human diseases. However many recent studies suggested that amyloids can also perform native function of the host organism, called functional amyloid. Many organisms including bacteria and fungi use these highly ordered amyloid fibrils for their adhesion and colonization to the surfaces. Recently we report that mammalian cells can also adhere on amyloids irrespective of sequence and absence of integrin recognition motif. This raises exciting possibility that amyloids could be primitive extracellular matrix. Many of our recent study also show amyloid based hydrogel is useful for designing smart biomaterials for tissue engineering application and drug delivery.

Talk: 3

Name of the Speaker: Dr. Manas Kumar Santra, Scientist E, Cancer Biology Division, National Centre for Cell Science, Pune.

Time: 12:45 – 1:10pm

F-box protein FBXO31 protects from oncogenic transformation through activation of cell cycle check points

Normal cells transformed into malignant cells either due to gain of function of proto-oncogene or loss of function of tumor suppressor gene or combination of both. Activation of oncogene leads to breaching the checkpoint activation barriers of malignant transformation. Through an RNAi screen we identified the F-box protein FBXO31, a candidate tumor suppressor encoded in 16q24.3 for which there is loss of heterozygosity in various solid tumors, as one of important factors required for oncogenic BRAF to induce senescence in primary human cells. We show that following oncogene-induced genotoxic stress, FBXO31 is phosphorylated by the DNA damage serine/ threonine kinase ATM, resulting in increased levels of FBXO31. FBXO31 then interacts with and directs the degradation of MDM2, which is dependent on phosphorylation of MDM2 by ATM. FBXO31- mediated loss of MDM2 leads to elevated levels of p53, resulting in growth arrest. In cells depleted of FBXO31, MDM2 is not degraded and p53 levels do not increase following stresses. Thus, FBXO31 is an essential checkpoint protein to prevent oncogenic transformation through activation of classic robust increase in p53 levels.

Session – III Talk: 1

Name of the Speaker: Dr. Ruchi Anand, Associate Professor, Department of Chemistry, Indian Institute of Technology (IIT), Bombay.

Time: 2:00 – 2:30pm

Structure Based Design of Aromatic Biosensors for Water Quality Monitoring

Phenol and its derivatives are common pollutants that are present in industrial discharge and are major xenobiotics that lead to water pollution. To monitor as well as improve water quality, attempts have been made in the past to engineer bacterial invivo biosensors. However, due to paucity of structural information there is insufficiency in gauging the factors that lead to high sensitivity and selectivity, thereby, impeding development. Here, we present the crystal structure of the sensor domain of MopR (MopRAB) from Acinetobacter calcoaceticus in complex with phenol and its derivatives to a maximum resolution of 2.55 Å. The structure reveals that the N- terminal residues 21-47 possess a unique fold, which are involved in stabilization of the biological dimer and the central ligand binding domain belongs to the 'nitric oxide signaling and golgi transport' fold, commonly present in eukaryotic proteins that bind long-chain fatty acids. In addition, MopRAB nests a zinc atom within a novel zinc binding motif, crucial for maintaining structural integrity. We propose that this motif is crucial for orchestrated motions associated with the formation of the effector binding pocket. Our studies reveal that residues W134 and H106 play an important role in ligand binding and are the key selectivity determinants. Furthermore, comparative analysis of MopR with XylR and DmpR sensor domains enabled design of MopR binding pocket that is competent in binding DmpR-specific ligands. Collectively, these findings pave way towards development of specific/broad based biosensors, which can act as useful tools for detection of this class of pollutants. Talk: 2

Name of the Speaker: Dr. Anuj Tripathi, Scientist, Nuclear Agriculture and Biotechnology Department, Bhabha Atomic Research Centre (BARC), Mumbai.

Time: 2:30 – 3:00pm

Macroporous Polymeric Cryogels: A Versatile System for Biomedical and Biotechnological Applications

Porous polymeric matrices are the key substrate in many bioengineering applications. In our recent finding we have developed and optimized thermally controlled cryotropic polymerization process for the synthesis of porous matrices. This method is an integrated one-step synthesis approach wherein the formations of porozen at low temperature as well as free-radical polymerization either chemically or by irradiation take place simultaneously under controlled conditions. The process parameters like degree of crosslinking, temperature, monomer’s concentration, physical/chemical moulding, volume and incubation time are important that provide a suitable environment to fabricate an ideal macroporous cryogel. Our approach is one of the simplest ways for the preparation of controlled porous scaffold with several other advantages like spongy nature, interconnected pores, mechanical stability, high solute flow and easy micro- sized particles mobility. In our preliminary study, physicochemical characterization suggests that wide range of modulation is possible in the properties of matrix like porosity, permeability, density, rigidity and elasticity, and also surface chemistry by tethering of functional moieties. The transitional changes between dry and wet state did not show change in its physico-chemical properties, which describe long term storage stability of these cryogels in dry as well as wet state. Vitally, this is a rapid method of synthesis of porous matrix with desired properties. We are currently exploring the potential of these cryogels for biomolecules/drug delivery, tissue- engineering, bioprocessing and environmental applications.

Keywords: Polymers, porous scaffolds, radiation, cryotropic, biomedical. Talk: 3

Name of the Speaker: Dr. Dr. Mayuri Gandhi, Research Scientist, Centre for Research in Nano Technology & Science (CRNTS), Sophisticated Analytical Instrument Facility (SAIF), Indian Institute of Technology (IIT), Bombay.

Time: 3:00 – 3:30pm

Session – IV

Talk: 1

Name of the Speaker: Dr. Prasanna Venkatraman, ACTREC

Time: 3:45 – 4:15pm

ATP binding sites and ATP hydrolyzing activity – surprise from studies on 14-3-3 proteins

14-3-3 proteins are a family of highly conserved proteins across eukaryotes. Seven known isoforms of 14-3-3 proteins have been identified in humans. 14-3-3 proteins exist as homo- and hetero-dimers and bind their target proteins at specific phosphoserine or phosphothreonine motif. They were originally identified as activators of neurotransmitter synthesis and subsequently as key regulatory proteins interacting with and modulating the activity of over 200 proteins involved in cell metabolism, signal transduction, cell cycle control, protein trafficking and stress responses. We identified a novel intrinsic ATP ase activity in this class of proteins. Residues that enhance or decrease this activity were identified by mutagenesis. Implications of this novel enzyme activity, stoichiometry of binding, the role of protein conformation and the puzzling details of conservation of this function or lack of it will be described. Talk: 2

Name of the Speaker: Dr. Subhash Bihani, Bhabha Atomic Research Centre (BARC), Mumbai.

Time: 4:15 – 4:35pm

Deinococcus FrnE: a novel cytoplasmic disulfide oxidoreductase

Disulfide oxidoreductases are ubiquitous redox enzymes that catalyze dithiol-disulfide exchange reactions in the cell. Trx (thioredoxin) and Grx (glutaredoxin) are two such enzyme systems that keep the free cysteine groups of intracellular proteins in a reduced state. These enzymes are members of Trx superfamily that is characterized by the presence of Trx fold and an active site CxxC motif. FrnE from Deinococcus radiodurans, a member of the Trx superfamily is induced on exposure of Deinococcus to Cd(II) stress and displays both disulphide reductase and isomerase activities. The crystal structures of recombinant FrnE protein show a typical thioredoxin domain with 22-CxxC-25 catalytic motif, a 7-helices insertion domain and unique C- terminal tail harbouring a novel 239-CxxxxC-244 motif. While in the native protein, FrnE shows active site (C22-C25) and C-tail (C239-C244) disulfide bonds, FrnE in the presence of DTT show an intermolecular disulphide bond between Cys-22 and Cys-244 residues. In presence of TCEP, FrnE shows a completely reduced active site. The interactions of the C-tail cysteines with active site cysteines suggest important regulatory/functional role of C-tail in the catalytic activities of FrnE. “True” FrnE orthologs, harbouring unique C-tail could be identified in genomes of seven bacterial phyla. Bioinformatics analysis suggested with high confidence cytoplasmic localization for FrnE orthologs. This is further confirmed by fluorescence microscopy studies of FrnE-GFP fusion protein in Deinococcus. We propose that FrnE represents a new cytoplasmic disulfide oxidoreductase system in prokaryotes.

PDB IDs: 5CNW, 5CO3, 5E59 References: Panicker L, Misra HS & Bihani SC. 2014. Purification, crystallization, and preliminary crystallographic investigation of FrnE, a disulfide oxidoreductase from Deinococcus radiodurans. Acta Crystallogr F70: 1540-1542. Talk: 3

Name of the Speaker: Dr. H. Muthurajan, Associate Professor, National Center for Nano Science and Nano Technology, Mumbai of University, Mumbai.

Time: 4:35 – 4:55pm

Packaging of Dry Electrodes for Bio-potential measurement and their Clinical Testing Standards

The bioelectrical signals carry useful information for understanding and diagnosing various pathological conditions of the patient. The origin of these bioelectrical signals is the change in the potential of the cells of the body tissues, giving rise to an electric field which transmits from one cell/neuron to another. These signals travel in the living epidermal (LE) layer which lies just below the stratum corneum (SC) or the outer most layer of the skin.

There exist a strong need and interest for EEG monitoring for medical conditions such as sleep apnea, epilepsy and traumatic brain injury. Thus, novel applications and uses are even more critical for the success of dry electrode technology for EEG. Thus, it is expected that these wireless, outpatient EEG-based neural monitoring systems will become much more common place in the near future. A robust and patient-friendly dry electrode system will be a significant contribution to this field.

The gel used along with gel based electrodes may also be associated withitching, allergic reactions and other skin problems in some patients. This method works well for short term monitoring, but subsequently the gel dries causing disruption in the signals. Despite several advances in the signal acquisition technology, the major obstacle which still remains is the transmission of signal at the skin-electrode interface. Dry electrodes or the gel free electrodes is another alternative method being widely explored in the field of healthcare devices.

We have successfully developed the packaging, sockets for EEG as well as ECG dry electrodes. This talk focus on necessity of biopotential measurement for patients as well as air-force operations and details on our research work for the same Talk: 4

Name of the Speaker: Dr. Susan Thomas, Scientist, National Institute of Research on Reproductive Health (ICMR), Mumbai.

Time: 4:55 – 5:15pm

Design of modulators of human follicle stimulating hormone receptor

The interaction of follicle stimulating hormone (FSH) with its receptor (FSHR) is crucial for reproductive health. Hence, molecules that can mimic this interaction can serve as fertility regulating agents. Structural analysis of the hormone-receptor complex has helped in identification of residues that confer receptor-binding specificity. Based on these residues and using in silico and wet-lab methods, few peptides and small molecules have been identified as potential FSHR modulators. The work related to the peptides will be discussed in brief during the presentation.