Training Data Analysis

SUMA SO MASEKH AR A N

* * **********, **** *** Jersey City, NJ 07305 2-03-540-**** s acee0r@r.postjobfree.com

Protein Biochemist with expertise in Molecular Biology, Cell Biology, Protein Engineering and medium

throughput Cell-based Assays with strong background in Structure-Function relationships of

Membrane Proteins and Epithelial Cell Biology. Prior training and experience in Clinical Biochemistry

and Diagnostic Assays. Seeking a research focused scientist position in a pharmaceutical/biotech

setting.

Core expertise in Molecular Biology, Protein Biochemistry, Protein Engineering and Cell Biology techniques.

Extensive experience in conducting medium throughput Cell-based Assays, Transport and Drug Inhibition

Kinetics, Biochemical Assays and Electrophysiology techniques.

Extensive experience in mammalian, insect and bacterial Cell Culture and Protein Expression in a variety of

cell lines.

Strong background in Structural Biology of Membrane Proteins and Biophysical and Computational tools

used in structural studies.

Strong experience in characterizing, and elucidating Mechanism of Action of Small Molecule Drugs.

Strong background in Epithelial Cell Biology, Epithelial and specifically Renal Transport Mechanisms.

Prior training in Clinical Biochemistry and Diagnostic Assays for metabolic diseases.

Proven track record in planning and executing large research projects. Very highly skilled in Experimental

Design and Data Analysis.

Extensive experience in lab management and mentorship . Strong verbal and written communication skills

with good p ublication record and extensive experience p resenting to expert audiences.

EXPERTISE

Protein Engineering: Expertise in using various Mutagenesis strategies (Model Directed Mutagenesis,

Substituted Cysteine Scanning, Tryptophan and Alanine Scanning)

Protein Function, Cell Based and Biochemical Assays (Medium throughput Cell-based Assays (96-well) –

assays for transport activity, kinetics and drug binding (Km, Vmax, EC50, IC50 ), Electrophysiology Assays-2

Electrode Voltage Clamp and Short Circuit Current Measurements, probing Disulfide Chemistry and Cysteine

Crosslinking Assays), Cell Viability and Counting, Oxidative Stress Assays, Enzymatic Assays (Protein Kinases,

Superoxide Dismutase), ELISA, Enzyme Kinetics, Metabolic Labeling using radiolabeled aminoacids and Pulse

Chase, Protein Quantification, Flow Cytometry (with formal training), ATP Assays. Extensive experience with

96-well Microplate Assays using high-end plate readers (Fluorescence, Luminescence, Colorimetry and

Radiometric). Very comfortable working with radioisotopes.

Diagnostic Biochemistry Assays: Tumor Marker Analysis, Serum Protein Analysis and Lipid Analysis. Cardiac,

Kidney and Liver Function Tests.

Protein Purification ; Immunoprecipitation, Affinity Purification, Size Exclusion Chromatography and HPLC

Protein characterization and biophysical Techniques: Extensive experience with Western blotting (using

Odyssey Infrared Imaging), Fluorescence Spectroscopy FRET, Circular Dichroism and NMR, 2-D Gel

Electrophoresis and Mass Spectrometry. Experience in using computational tools such as Visual Molecular

Dynamics (VMD) and Molecular Operating Environment (MOE) and Modeller.

Molecular Biology: PCR, RT-PCR, Site -directed Mutagenesis, Molecular Cloning; designing and making

constructs (including insertions and deletions), Baculovirus Cloning, RNA and DNA extraction, mRNA and

cDNA Synthesis, Primer Design, Restriction Digest Analysis and DNA Sequencing Analysis. Adept in using

Lasergene.

Cell Biology: I mmunohistochemistry including labeling of Extracellular Epitopes, Live Cell Imaging using

Confocal Microscopy (Zeiss 710).

Cell Culture and protein expression : Extensive experience with Mammalian Cell Culture (cell lines from

kidney, gut and vasdefrens-HEK, CHO, HeLa, MDCK, T-84, IPEC-J2, CACO-2, PVD9902 and Primary Cell Culture),

human red blood cells, insect cells (SF9), bacterial cell culture and modest experience working with rats and

rat tissue and frog dissections. Extensive experience in mammalian cell transfections and generating stable

cell lines. Protein expression in bacterial and insect cells.

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Data Analysis: Excel and Sigma Plot, Curve fitting (Transport Kinetics, Michaelis Menten Kinetics, Hill Fits,

Sigmoidal and Exponential Fits and Deriving Hill Coefficients, Time and Rate Constants.), Adobe

Illustrator/Photoshop, Corel Draw, ImageQuant, and Statistical Analysis (ANOVA and t-tests).

Other Skills and Experience:

Presentation : Experience presenting at several national conferences including FASEB, Annual Biophysical

Society, Annual Cell Biology meetings and Gordon Conferences. Presenting at inter and intradepartmental

meetings.

Leadership : Laboratory management, ordering lab equipment and managing repairs, coordinating and

managing compliance with environmental health and safety. Mentored undergraduate and new graduate

students.

Teaching: Extensive teaching experience – 2 years teaching undergraduate senior level Biochemistry lab and

one semester teaching graduate level Protein Structure lab.

Coursework: Extensive coursework in all aspects of protein structure: Protein structure, Physical

Biochemistry, Protein Modeling NMR, Protein Structure and NMR lab, Protein Modeling Lab, MOE Training,

Nanocourse in conducting high throughput assays at the Yale Center for Molecular Discovery and Courses in

Flow Cytometry (3 day comprehensive and Invitrogen Online tutorial).

SUMMARY OF P ROJECTS

YALE UNIVERSITY, New Haven, CT Postdoctoral Associate 11/08 to 01/14

Department of Cellular & Molecular Physiology

Conducted structure function studies of transmembrane domains of sodium-potassium chloride co-

transporter (NKCC1) using site directed mutagenesis, substituted cysteine and tryptophan scanning, MTS

accessibility studies, crosslinking studies and functional assays (transport and drug binding kinetics) to

validate a homology model of NKCC, identify key ion binding and loop diuretic binding residues and study

mechanism of activation of NKCC1. Engineered partially and fully cysless NKCC.

Studied membrane expression and localization of NKCC1 engineered to have extracellular epitope tags and

protease sites.

Performed cloning, expression and purification of C terminus of NKCC1: using PET and Fast Bac vectors and

transfection, expression and protein purification of NKCC1 C-terminus from SF9 (insect) cells.

YALE UNIVERSITY, New Haven, CT Short-term Research Position 02/08 to 11/08

Studied protein-protein interactions using Tandem Affinity Purification (TAP) of Serum Glucocorticoid Kinase

(SGK).

KANSAS STATE UNIVERSITY, Manhattan KS Graduate Research Assistant 01/02 to 12/07

Position held while pursuing PhD in Biochemistry

Doctoral project: Characterization and studying mechanism of action of channel forming peptides designed to

increase transepithelial chloride transport and paracellular drug transport.

Studied reversible reorganization of epithelial tight junctions by channel forming peptides.

Studied the signaling pathways involved in the reorganization of tight junction proteins upon exposure to

different modulators of barrier function.

NATIONAL CENTRE FOR BIOLOGICAL SCIENCES, Bangalore, India Research Training 10/00 to 05/01

Cognitive experiments on male and female rats subject to chronic restrain stress.

KASTURBA MEDICAL COLLEGE, Manipal, India Graduate Research Assistant 09/97 to 06/00

Position held while pursuing MS in Clinical Biochemistry

Designed Clinical study to study oxidative stress in premenstrual syndrome. Conducted biochemical assays to

study antioxidant levels (superoxide dismutase, Vitamin C and E) and lipid peroxidation in RBCs.

I nternship in a diagnostic laboratory: Tumor marker assays, serum lipid analysis, biochemical assays for

cardiac, liver and kidney function tests

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EDUCATION

Ph.D. in Biochemistry

Kansas State University, Manhattan, KS (09/01 to 12/07)

MS in Clinical Biochemistry

Kasturba Medical College, Manipal, India (06/00)

Human Anatomy, Human Physiology, Biochemistry & Human Clinical Biochemistry with training in the Clinical

Biochemistry Laboratory

Bachelor of Science in Chemistry, Botany and Microbiology

Bangalore University, Bangalore, India (05/97)

P UBLICATIONS

Somasekharan S, Monette M, Forbush B. Homodimerization of NKCC1 at the interface of transmembrane

domains 11 and 12, in preparation.

Monette M, Somasekharan S, Forbush B. Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion

Transport and Transporter Activation Revealed by Internal Cross-linking between Transmembrane Domains

10 and 11/12 J Biol Chem. 2014 Mar 14;289(11):7569-79.

Somasekharan S, Monette M, Forbush B. Functional Expression of Human NKCC1 from a Synthetic Cassette-

Based cDNA: Introduction of Extracellular Epitope Tags and Removal of Cysteines, Dec 5 2013, PLoS ONE.

Suma Somasekharan, Jessica Tanis and Biff Forbush Loop diuretic and ion-binding residues revealed by

scanning mutagenesis of transmembrane helix 3 (TM3) of Na-K-Cl cotransporter (NKCC1), May 18, 2012 The

Journal of Biological Chemistry, 287, 173**-*****.

Migliati E, Meurice N, DuBois P, Fang JS, Somasekharan S, Beckett E, Flynn G, Yool AJ I nhibition of

aquaporin-1 and aquaporin-4 water permeability by a derivative of the loop diuretic bumetanide acting at an

internal pore-occluding binding site., Mol Pharmacol, 2009 Jul;76(1):105-12.

Somasekharan S, Brandt R, Iwamoto T, Tomich J M, Schultz BD. Epithelial barrier modulation by a channel

forming peptide., J Membr Biol, 2008 Mar; 222(1):17-30.

Gargi Kalia, Suma Sudheendran (co –author) and Anjali Rao Antioxidant status and lipid peroxidation in

premenstrual syndrome: a preliminary study, Clin Chim Acta 2001 Jul 5; 309(1):97-9 (Please note: This was a

co- authorship based on research done for my master’s thesis. There is an editorial error in my last name.)

AWARDS

2007: Phi Zeta Podium Presentation Award

2006: Terry. C. Johnson Summer Research Award

2006: American Society of Cell Biology Travel Award

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