We like to thank the Department of Biotechnology (Government of India) for financial support. We like to thank our sponsors for financial support: Platinum Sponsor: Gold Sponsor: Silver Sponsors: And Program Date Venue Timings Title OF MUSIC, MATH AND MEASUREMENT LH1 6pm M.W. Linscheid, Humbold Universitaet Berlin, Germany Dining Mixer 23/08 7pm Complex Concert by Ananth Menon Quartet Dining 8pm Dinner Complex LH1 9:00am Tutorial I MASS SPECTROMETRIC ESSENTIALS IN OMICS RESEARCH D.Schwudke LH1 9:30am Session I / Proteomics (Chair D. Schwud ke) MASS SPECTROMETRIC IDENTIFICATION OF A NOVEL TRANS- SPLICING EVENT IN GIARDIA LAMBLIA HEAT SHOCK PROTEIN 90 U.S. Tatu, IISC Bangalore, India REVISITING PROTEOMICS OF GLIOMAS: DIFFERENTIAL MEMBRANE PROTEINS AND MOLECULAR INSIGHTS R. Sirdeshmukh, IOB Bangalore, India NEW HIGH SELECTIVITY WORKFLOWS FOR TARGETED QUANTITATIVE PROTEOMICS 24/08 M. Cafazzo, AB Sciex, US Break ABSOLUTE QUANTIFICATION OF PROTEINS AND PEPTIDES - USE OF METAL CODING AND LC/MS WITH ICP AND ELECTROSPRAY M.W. Linscheid, Humbold Universitaet Berlin, Germany MATERNAL VITAMIN B12 DEFICIENCY INDUCED ALTERATION IN PROTEIN EXPRESSION IN RAT OFFSPRING S. Sengupta, IGIB New Dehli, India CLINICAL PROTEOMICS OF EYE DISEASES K. Dharmalingam, Madurai Kamaraj University, India Dining 1pm Lunch Complex Ope n 2pm Poster Session I Deck Date Venue Timings Title LH1 4pm Session II / Lipidomics (Chair S. Hebbar ) TOWARDS THE COMPLETE STRUCTURE ELUCIDATION OF COMPLEX LIPIDS BY MASS SPECTROMETRY: NOVEL APPROACHES TO ION ACTIVATION S. Blanksby et al., University of Wollongong, Australia LIPIDOMICS AT THE HIGH MASS RESOLUTION A. Shevchenko, MPI-CBG Dresden, Germany STRATEGIES FOR IMAGING BIOMOLECULES BY TOF-SIMS AND MALDI-TOF/TOF MASS SPECTROMETRY O. Laprévote, Université Paris Descartes, France Break 24/08 MEIBUM LIPID COMPOSITION IN ASIANS WITH DRY EYE SYNDROME M.R. Wenk, NUS, Singapore INTEGRATION OF OMICS RESEARCH WITH DEVELOPMENTAL BIOLOGY D. Schwudke, NCBS Bangalore, India ADIPOSE TISSUE LIPIDOME: BENEFITS AND COSTS OF LIPID REMODELING AS ADAPTATION TO ACQUIRED OBESITY M. Orešič, VTT Technical Research Centre of Finland NN D. Ghosh, JNU Dehli, India Dining 8pm Dinner Complex Date Venue Timings Title LH1 9:00am Tutorial II SOFT IONIZATION METHODS K. Dreisewerd LH1 9:30am Session III / Proteins and Proteomics (Chair A. Shevchenko) GAS PHASE STRUCTURAL AND DYNAMICAL BIOLOGY J.L. Benesch, University of Oxford, UK IDENTIFICATION OF MULTIPLE FOLDING PATHWAYS OF MONELLIN USING PULSED THIOL LABELING AND MASS SPECTROMETRY J.B. Udgaonkar et al., NCBS Bangalore, India DIFFERENTIAL EXPRESSION OF RED CELL PROTEINS IN HEMOGLOBINOPATHY A. Chakrabar , SINP Kolkata, India 25/08 Break COMPARATIVE PROTEOMICS OF EXTRACELLULAR MATRIX DEMONSTARTES THE COORDINATED EXPRESSION OF DEHYDRATION-RESPONSIVE PROTEINS N. Chakraborty, NIPGR Dehli, India ON-LINE UPLC/ION MOBILITY SEPARATION/TOF MS FOR QUALITATIVE AND QUANTITATIVE PROTEIN PROFILING IN COMPLEX BIOLOGICAL SAMPLES M.A. McDowall et al., Waters, Manchester, UK A DRAFT MAP OF THE HUMAN PROTEOME H. Gowda, IOB Bangalore, India Dining 1pm Lunch Complex Ope n 2pm Poster Session II Deck Date Venue Timings Title LH1 4pm Session IV / Metabolites and Small Molecules (Chair R. Kanna n) G-QUADRUPLEX FORMATION OF GUANOSINE DERIVATIVES IN PRESENCE OF ALKALINE EARTH METAL IONS STUDIED BY ELECTROSPRAY IONIZATION MASS SPECTROMETRY M. Vairamani, IICT Hyderabad, India IR-MALDI MASS SPECTROMETRY FOR THE ANALYSIS OF (SMALL) MOLECULES DIRECTLY FROM BIOLOGICAL TISSUE AND FROZEN AQUEOUS SOLUTIONS K. Dreisewerd et al., University of Münster, Germany QUANTITATIVE ANALYSIS OF SMALL BIOMOLECULES USING LASER DESORPTION IONIZATION MASS SPECTROMETRY V. Panchagnula, NCL Pune, India DISCOVERY OF NOVEL DROSOPHILA LIPID PHEROMONES USING MASS SPECTROMETRY 25/08 J. Yew et al., Temasek Life Sciences Laboratory, Singapore Break MULTI-OMICS WORKFLOW FOR STUDYING DRUG TREATMENT ON HUMAN CELL LINE S. Rajagopalan, Agilent Technologies, India LIQUID EXTRACTION SURFACE ANALYSIS (LESA) COMBINED WITH NANOESI-MS FOR DIRECT SAMPLING OF SURFACES R. Almeida, Advion BioSciences, UK NOVEL APPROACHES BASED ON HIGH RESOLUTION AND ION MOBILITY MASS SPECTROMETRY FOR THE QUAL/QUAN ANALYSIS OF PHARMACEUTICALS AND THEIR METABOLITES G. Hopfgartner, Université de Genève, Switzerland Dining 8:00pm Dinner Complex LH1 9:30am Short Talk Session (Students Presenta ons) (Chair Schwudke Lab) 26/08 LH1 11:00am Interac on LH1 12:00am Poster Prizes - Cer ficates LH1 12:30am Closing Remarks Speakers Abstracts LC/MS with iFunnel Technology gives you the lowest limits of detection—down to zeptomole levels at conventional flow rates. GC/MS/MS acquisition speed up to 500 MRM transitions per second lets you automatically quantify and confirm more targets in a single method. ICP-MS High Matrix Introduction (HMI) technology enables faster, easier ppt-level quantification of trace elements in the dirtiest samples. CE/MS is the industry’s only single-vendor, fully integrated solution delivering flexibility, performance, and productivity—plus the confidence of Agilent reliability. Agilent MassHunter Software makes your MS analyses faster, easier and more productive with software that is consistent for all your MS instruments. CHECK OUT ALL THE REASONS WE’RE THE MASS SPEC LEADER: www.agilent.com/chem/clearlybetter © Agilent Technologies, Inc. 2011 24/08 – Tutorial I MASS SPECTROMETRIC ESSENTIALS IN OMICS RESEARCH Dominik Schwudke National Centre for Biological Sciences, TIFR, Bangalore / Bengaluru, INDIA [email protected] Many mass spectrometric experiments performed in context of biological research are belonging to the Omics domain. I will comment on the design of high-throughput workflows to measure entire pools of biomolecule classes. In that context I will give an overview on how mass spectrometric parameters like resolution and mass accuracy influence our ability to interpret OMICS data in qualitative and quantitative manner. We will discuss terms like dynamic range, mass resolution and separation power of a mass spectrometric approach. Further we will discuss the ability to decrease false positive and false negative assignments by using accurate masses and isotopic distributions. 24/08 Session I – Proteomics Talk 1 MASS SPECTROMETRIC IDENTIFICATION OF A NOVEL TRANS-SPLICING EVENT IN GIARDIA LAMBLIA HEAT SHOCK PROTEIN 90 Utpal Tatu Professor, Department of Biochemistry, Indian Institute of Science, Bangalore, India [email protected] Material and Methods My laboratory is interested in examining in vivo roles of molecular chaperones in protozoa, many of which cause infectious diseases in humans and animals. We have examined Hsp90 gene expression and function from Dictyostelium discoideum, Plasmodium falciparum, Trypanosoma evansi as well as Giardia lamblia. G. lamblia trophozoites are grown to confluency and total protein was extracted. Proteins were resolved in SDS PAGE. Band corresponding to Hsp90 was subjected to in-gel trypsinization and peptides were enriched. Peptides were separated in nano- reverse phase column which was directed to Nano Spray ESI- QTOF MS. Ionized peptides were analyzed by one full MS scan and four consecutive product ion scans of the four most intense peaks. The acquired data was analyzed using ProteinPilot 4.0 software with combined Giardia database. Peptide confidence greater than 90% was considered as significant hits. Spectra of each peptide used for identification of the proteins were verified manually. Results In my talk I will specifically describe our recent results incorporating mass spectrometric analysis of Hsp90 from G.lamblia, which revealed a novel trans-splicing based expression of GlHsp90 in this minimalist protozoan parasite. We have recently shown that Hsp90 in Giardia is arranged as a split gene, HspN and HspC separated by 777 kb intergenic sequence. The full length Hsp90 transcript is a resultant of a novel trans-splicing phenomenon which is mediated by spliceosomal complex. The mature full length transcript has all the hallmarks of the canonical Hsp90. The protein band that corresponded to a region of around 80 kDa was identified to be GlHsp90, with peptides matching from two individual exons, HspN and HspC. In addition, during the course of trans-splicing there is incorporation of 99 nts at the junction of two exons which was obtained by sequencing of full length cDNA. Since the Giardiadb database does not contain the full-length GlHsp90 sequence, we manually included the protein sequence in our search list by translating the cDNA sequence obtained. Using mass spectrometric based approach we have sequenced and identified the peptides joining these two exons. This junctional peptide houses an ‘Arg’, a catalytical residue conserved in other Hsp90s, required for the functioning of Hsp90 chaperone cycle. We have also identified ‘Val’, a conserved residue across Hsp90s, formed during the junction of the two ORFs. Thus the sequenced peptides scan the entire junction of the full length Hsp90. 24/08 Session I – Proteomics Talk 1 Conclusions Hsp90 is a conserved molecular chaperone required for normal functioning of the cell. Hsp90 also provides survival advantages at time of stress to the baring organisms. Hsp90s indentified so far is 80 – 90 kDa in size and encoded by a single ORF. Interestingly in Giardia Hsp90 is encoded by two different ORFs which can be functional only when brought together by a post transcriptional mechanism, trans-splicing. Here we provide evidence for such a reconstruction mechanism at proteomic level.