Project Research

JAGANATHAN LAKSHMANAN, Ph.D. (JAGAN)

****, ******** *****, **********, ** 40207. USA.

Tel: 502-***-****

e-mail: *******@*****.***

KEY STRENGTHS

Highly motivated, innovative, and performance driven scientist equipped with proven expertise in protein biochemistry and molecular cell biology

• Expertise in protein expression and purification from bacteria, yeast, insect and other systems.

• Extensive experienced in FPLC, HPLC, SDS-PAGE, IEF and CD and fluorescence spectroscopy

• Extensive experience in sample preparation and data analysis for MS, MALDI-TOF MS

• Working knowledge on cGLP/cGMP

• Excellent project management and technical leadership skills, successfully trained and managed scientific staff

• Proven track record of successful research with 13 research papers published in high-quality peer reviewed journals

• Earned PhD in Biochemistry with honors (magna cum laude)

• Extremely adaptable, solid interpersonal skills and worked collaboratively with people of diverse cultural backgrounds

• Experience working in cross-functional team environment and excellent problem solving abilities

KEY SKILLS / ATTRIBUTES

Protein expression & purification

AKTA FPLC & HPLC

UV&CD Spectroscopy

Protein PAGE (Native & SDS)

SDS-PAGE, IEF, 2D-DIGE

Filtration methods

Assay/methodology development

Protein refolding

Data analysis and interpretation

Analysis and problem resolution

Time management

Strong work ethic

WORK ELIGIBILITY

Permanent resident (Green card holder)

EDUCATION

Ph.D. Biochemistry, Bharathiar University, Coimbatore, India, 2001, graduated with Magna cum laude

M.S. Biochemistry, Bharathiar University, Coimbatore, India, 1993

B.S. Biochemistry, Bharathiar University, Coimbatore, India, 1991

RESEARCH AND PROFESSIONAL EXPERIENCE

a) Research Faculty: Psychiatry/Oral Health/Surgery 2007-present, University of Louisville, KY, USA.

• Purified and characterized novel isoforms of inositol -1-phosphate-synthase (Isyn) enzyme from rat tissues.

• Expressed and purified human Isyn enzyme for antibody generation. Designed antigen to target phosphorylated form of Isyn enzyme.

• Characterized brain inositol signaling pathway to identify the pathway’s role in neuro-psychiatric disorders like Bipolar Disorder and Schizophrenia.

• Identified critical transcription factors involved in the development of rat parotid gland.

• Elucidated the role of Gsk3beta in IL-1beta mediated up-regulation of hepatic iNOS.

Techniques employed: Mass Spectroscopy, SDS-PAGE, western blot, EMSA and 2D-DIGE, UV/Visible & fluorescence spectroscopy, CD spectroscopy, immuno-histochemistry, confocal fluorescence microscopy, cell culture (primary and stable cell lines), animal handling and surgery.

Departmental Responsibility: Supervise, provide support and training for research fellow, graduate students and lab technicians; analysis of data and preparation of manuscripts for submission to journals; presentation of research results in conferences and symposiums; supporting the team-leader in the preparation and submission of grant proposals to funding agencies.

Funded Grants:

• Co-Investigator, Mood-stabilizing medications and the inositol signaling system. 2010-2014. VA Merit Review Grant. Project No.1I01BX000387-01A2

• Co-Investigator, “Double Blind, Placebo-Controlled, Crossover Study of Glutathione Vitamin C and Cysteine in Children with Autism and Severe Behavior Problems. 2008-present. Kosair Children’s Hospital Foundation. HSPPO #08.0220.

b) Post-doctoral Research Scholar:Molecular Neuroscience Laboratory/Brown

Cancer Center,2003-2007, University of

Louisville, Louisville, KY, USA.

• Cloned and over-expressed the transmembrane GPCR, CXCR4 in yeast and mammalian cells.

• Expressed the chemokine SDF-1 in bacterial system and characterized the protein by MS and NMR.

• Developed cell based system for identifying against /antagonist for chemokine receptor CXCR4.

• Identified and characterized novel isoform and phosphorylated form of inositol synthesizing enzyme employing MS/MALDI-TOF.

• New targets of lithium were discovered by 2D-DIGE and MS proteomic analysis of rat prefrontal cortex.

• 3 publications in peer-reviewed journals and another 4 manuscripts under preparation; co-investigator on one major research grant funded by Veterans Affairs Merit Review.

Techniques employed: Mass Spectroscopy, HPLC/FPLC, SDS-PAGE, western blot and 2D-DIGE, Fluorophore-assisted Carbohydrate Electrophoresis analysis of glycans UV/Visible & fluorescence spectroscopy, CD spectroscopy, immunohistochemistry, confocal fluorescence microscopy, mammalian cell culture (primary and stable cell lines), animal handling and surgery.

Departmental Responsibility: Supervise, provide support and training for graduate students and lab technicians.

c)Post-doctoral scholar: Molecular and Cellular Biochemistry Department,

2001-2003,University of Kentucky, Lexington, KY, USA.

• Discovered the phosphorylated form the yeast transcription factor Rgt1 and its involvement in regulation of yeast glucose transporter gene expression

• cDNA library form MIN6 cell line for use in yeast 2-hydrid system

• 3 publications in peer-reviewed journals and 1 methodology article in Methods in Molecular Medicine.

Techniques employed: Mammalian cell culture, adenoviral based transfections, molecular biology techniques, yeast and bacterial transformations, protein SDS-PAGE and western-blotting, affinity chromatography, immunohistochemistry, fluorescence microscopy

Post-doctoral Fellowship – American Heart Association (Ohio Valley Affiliate, USA): Project reference No.: 0325322B (07/01/2003). Title of Project: Analysis of the role of O-linked GlcNAc transferase (OGT1) in the regulation of insulin gene transcription.

d) Research Associate: Biochemistry, 2000-2001

L. V. Prasad Eye Institute, Hyderabad, India.

• Purification of eye-lens crystallins from bovine and human lens.

• Analysis of cataract lens proteome using 2D-PAGE and MS

• Identified that alpha-crystallins do not lose its chaperon activity even after structural modification by peroxinitrite free radical.

Techniques employed: Mass Spectroscopy, HPLC, AKTA FPLC, UV-visible spectroscopy, CD and fluorescence spectroscopy, 2D-protein gel, column chromatography for protein purification.

Departmental Responsibility: Negotiate and order instruments and supplies for the lab, train graduate students about GLP and research methodologies.

e)Doctoral Scholar: Department of Biotechnology, 1994-2000

Bharathiar University, Coimbatore, India.

• Purification and characterization of cholinesterase enzyme form human plasma.

• Developed a method to stain the aryl acylamidase activity of cholinesterases after native-PAGE.

• Thermo-stabilized human serum cholinesterase and tested its suitability for the detection of organophosphates and carbamates in biological and environmental samples.

• Purification and characterization of lipase form fungi.

Techniques employed: Ion-exchange/affinity/size-exclusion chromatography, chemical coupling of affinity ligands to Sepharose matrix, enzyme assay and kinetics, SDS-PAGE, UV/visible spectroscopy, fluorescence spectroscopy

Departmental Responsibility: Provide support and training for protein purification techniques for new graduate students of the lab; manage lab instruments.

RESEARCH PUBLICATIONS

1. Seelan RS, Lakshmanan J, Casanova MF and Parthasarathy RN. Identification of myo-inositol 3-phosphate synthase isoforms: Characterization, expression and putative role of a 16 kDa gamma c isoform. Journal of Biological Chemistry, 28, 9443-9457 (2009).

2. Seelan RS, Khalyfa A, Lakshmanan J, Casanova MF and Parthasarathy RN. Deciphering the lithium transcriptome: microarray profiling of lithium modulated gene expression in human neuronal cells, Neuroscience, 151, 1184-97 (2008).

3. Thiagarajan G, Lakshmanan J, Chalasani M, and Balasubramanian D. Peroxynitrite Reaction with eye lens proteins: crystallin retains its activity despite modification, Investigative Ophthalmology & Visual Science, 45, 2115-2121 (2004).

4. Ozcan S, Mosley AL, Lakshmanan J, and Aryal BK. Glucose regulation of HXT gene expression by the transcription factor Rgt1. Yeast, 20, S166-S166 Suppl. 1 (2003).

5. Lakshmanan J, Elmendorf JS and Ozcan S. Analysis of insulin-stimulated glucose uptake in differentiated 3T3-L1 adipocytes. Methods in Molecular Medicine, 83, 97-103 (2003).

6. Lakshmanan J, Mosley AL and Ozcan S. Repression of transcription by Rgt1 in the absence of glucose requires Std1 and Mth1. Current Genetics, 44, 19-25 (2003).

7. Mosley, A.L*, Lakshmanan J*, Aryal BK, and Ozcan S. Glucose-mediated phosphorylation converts the transcription factor Rgt1 from a repressor to an activator, Journal of Biological Chemistry, 278, 103**-***** (2003). * Equal contribution

8. Lakshmanan J, and Boopathy R. A direct method to visualize the aryl acylamidase activity on cholinesterases on polyacrylamide gels. BMC Biochemistry, 1, 3 (2000). http://www.biomedcentral.com/1471-2091/1/3

9. Lakshmanan J, and Boopathy R. Distinct Effect of Benzalkonium Chloride on the Esterase and Aryl Acylamidase Activities of Butyrylcholinesterase. Bioorganic Chemistry, 28, 242-251 (2000).

10. Lakshmanan J, and Booapthy R. Thermostabilization of Human Serum Butyrylcholinesterase for detection of its inhibitors in water and biological fluids. Brazilian Archives of Biology & Technology, 42, 343-348 (1999).

11. Ganguraj U, Lakshmanan J, Shridas P and Boopathy R, Purification and characterization of lipase from Rhizomucor miehei. Journal of Scientific & Industrial Research, 57, 607-610 (1998).

12. Lakshmanan J, and Boopathy R. Hydrophobicity on esterase activity of Human Serum Cholinesterase. Interaction of Triton X-100 with acyl pocket of butyrylcholinesterase: Effect on esterase activity and inhibitor sensitivity of the enzyme. Indian Journal of Biochemistry & Biophysics, 35, 142-147 (1998).

13. Lakshmanan J, Padmalatha K, Revathi G and Boopathy R, in “Enzymes of the Cholinesterase Family”, Eds., Quinn DM, Balasubramanian AS, Doctor BP and P.Taylor, Plenum press, New York, pp 123-124 (1995).

COMPUTER SKILLS

Analysis software: Mascot, ProteinProspector, Vector NTI, Graphpad, Prism, Igor pro, ImageJ, Ingenuity Pathway Analysis program.

Productivity: Microsoft Word, Excel, PowerPoint, Adobe Illustrator, Photoshop.

Operating systems: Mac OS 9 and X, Linux, Windows 95, 98, XP, Vista and Windows 7.

PROFESSIONAL AFFILIATIONS

American Association of Dental Research

Society of Neurosciences

American Association for Advancement of Science -2008

Indian Eye Research group -2001

Society of Biological Chemist, India -2000

ACADEMIC SERVICE

2009-present : Member – ACORP sub-committee, VA Medical Center, Louisville, KY

2000-2001 : Lecturer in Biochemistry – Bausch & Lomb School of Optometry, Kismathpur, Hyderabad, India.

PRESENTATIONS AT INTERNATIONAL CONFERENCES (Select)

1. Lakshmanan J, Adcock S, DeLeeuw L and Lane AN. “High level expression and purification of chemokines for structural studies”. Fourth Annual Peptalk – The Protein Information Meet, San Diego, USA, January 10-13, 2005.

2. Lakshmanan J, Trent JO and Lane NA. "Biophysical characterization of the CXCR4 receptor and interaction with ligands". 4th Annual Scientist's Seminar of Kentucky Lung Cancer Research Program, Frankfort, KY, USA, October 20, 2006.

3. Lakshmanan J, Seelan RS, Casanova MF and Parthasarathy RN. Role of phosphorylation in the regulation of inositol levels in the brain. Neuroscience 2007, Society of Neuroscience Meeting, San Diego, CA, USA, November 2-7, 2007 (Poster # Y13).

4. Lakshmanan J, Seelan RS, Casanova MF and Parthasarathy RN. Myo-Inositol 3-phosphate synthase (Isyn) Knockout (KO) mouse exhibits syndactyly and other physical abnormalities. 18th Annual Neuroscience Day Louisville Chapter of Society for Neuroscience, Louisville, KY, USA. April 17, 2008.

5. Lakshmanan J, Parthasarathy R, Seelan RS and MF.Casanova. Proteomic analysis of Isyn knockout mice prefrontal cortex. 19th Annual Neuroscience Day Louisville Chapter of Society for Neuroscience, Louisville, KY, USA. April 16, 2009.

6. Lakshmanan J, Seelan RS, MF.Casanova and Parthasarathy R. Proteomic and pathway analysis of rat prefrontal cortexafter chronic lithium treatment. Neuroscience 2009, Society of Neuroscience Meeting, Chicago, IL, USA, October 12-21, 2009 (Poster # 839.5/M12).

7. Lakshmanan J, Venkatesh SG, Carenbauer, Waige Sl, Zacharias W and Darling, DS. Xbp1 regulates Mist1 and PSP in parotid gland. International Association of Dental Research 2011 Annual Meeting, San Diego, CA, USA. March 16-19, 2011.

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