Vatsala Sagar

**** ********* ***** ***********, ** 20783

301-***-****

abq0u2@r.postjobfree.com

OBJECTIVE: My work experience spans Molecular Biology, Crystallography,

Protein Purification, and Immunology. I am looking forward to support an

organization with the development and analysis of their current line of

products and to assist in the research and development of new biomedical

products and services.

EDUCATION

Ph.D. in Biochemistry. 2007-2011

University of Maryland College Park. Department of Chemistry and

Biochemistry,

under Dr. Nicole LaRonde-LeBlanc

M.S. Biochemistry, Chemistry and Molecular Biology. 2001-2003

Johns Hopkins University School of Medicine,

under Dr. Phillip Cole, Director of Pharmacology and E.K. Marshall and

Thomas

H. Maren Professor

B.S. in Biochemistry and Molecular Biology. 1998-2001

University of Maryland Baltimore County

MAGNA CUM LAUDE recipient

SCIENTIFIC AUTHORSHIP- PUBLICATIONS

1. Ferreira-Cerca, S.*, Sagar, V.*, Schafer, T., Diop, M., Wesseling, A.,

Lu, H., Chai, E., Hurt, E., LaRonde-LeBlanc, N. (2012) ATPase-dependent

role of the atypical kinase Rio2 on the evolving pre-40S ribosomal

subunit, Nat Struct Mol Biol 19(12):1316-23.

2. Shi, D., Sagar, V., Jin, Z., Yu, X., Caldovic, L., Morizono, H.,

Allewell, N. M., and Tuchman, M. (2008) The crystal structure of N-acetyl-

L-glutamate synthase from Neisseria gonorrhoeae provides insights into

mechanisms of catalysis and regulation., J Biol Chem 283, 7176-7184.

3. Zheng, J.*, Sagar, V.*, Smolinsky, A., Bourke, C., LaRonde-LeBlanc, N.,

and Cropp, T. A. (2009) Structure and function of the macrolide biosensor

protein, MphR(A), with and without erythromycin., J Mol Biol 387, 1250-

1260.

4. Sagar, V., Zheng, W., Thompson, P. R., and Cole, P. A. (2004)

Bisubstrate analogue structure-activity relationships for p300 histone

acetyltransferase inhibitors., Bioorganic & medicinal chemistry 12, 3383-

3390.

* Indicates joint first authorship.

PROFESSIONAL EXPERIENCE

University of Maryland College Park, 2007-2011

Research and Graduate Research/Teaching Assistant

. Focused research to study enzymes not readily amenable to purification

and crystallization. I independently worked on identifying the

eukaryotic protein structure of ribosomal processing factor, Rio2, at

2.2 . My research on the structural analysis of Rio2 has uncovered a

new catalytic intermediate suggesting an alternate role to Rio2,

originally designation as a kinase. My groundbreaking research also

identified that the Rio2 acts as an ATPase switch to confer that small

ribosome maturation is complete. Over a three year period, I

successfully purified Rio2 from 40% close molecular weight degradation

products and crystals of Rio2 were enlarged from microcrystals

following a innovative 4 month process.

. Found the first crystal structure of N-acteylglutamate synthase (NAGS)

complexed with acetyl-CoA and with CoA plus N-acetylglutamate (NAG) at

2.5 and 2.6 . Impairment of NAGS results in blockage of ureagenesis

leading to hyperammonemia. NAGS has been exhaustively studied since

1981 but determination of a crystal structure had been difficult. In

2006 Shi, et. al wrote of negative results in realizing NAGS crystal

structure. In the first year of my PhD studies I unmasked how to

crystallize NAGS so that it diffracted to 2.4 . My structural

analysis revealed NAGS forms a hexamer with six active sites. A

monomer has two subunits A and B and a single active site consists of

subunit A from monomer 1 and subunit B from adjacent monomer 2. The

six monomers form a ring and activity is concomitant on the structure

of the ring being maintained.

. Collaborated on the structure of MphR(A), a macrolide biosensor

protein that acts as a repressor of transcription of the erythromycin

2'-phosphotransferase MphA gene. I determined the crystal structure

of MphR(A) in the apo form and complexed with erythromycin.

Understanding the mechanism of MphR(A) will lead to advancement in

combatting growing drug resistance of infectious bacteria.

. Worked independently or supervised teams of two on all projects.

I have skills in project design, cloning, protein purification,

crystallization, structure solution/ refinement and analysis. My work on

determining the crystal structures of these diverse and difficult to manage

proteins has given me knowledge and skills to solve most research

assignments. I intend to do research to become a successful scientist in

any Biological or Biochemical field. Concurrently, I have five years

teaching experience in Organic Chemistry and Analytical Chemistry

Laboratories and have assisted in General Biochemistry and General

Chemistry instructions.

Edge Biosystems Gaithersburg MD, 2005-2006

Research Associate in Research and Development

. Collaborated with senior scientists on the development of a new

product to purify DNA via the use of magnetic beads.

. Researched and developed a technique of my own to bind and

remove DNA from magnetite, an iron oxide. This allows for

purification of DNA from heterogeneous mixtures.

Antex Biologics, Inc. Gaithersburg MD, 2004-2005

Research Associate in Research and Development

. Worked as a collaborator with the senior scientist to design,

clone and purify constructs for possible antigens of the

chylamydia vaccine. Identified several soluble constructs for

vaccine studies.

Naval Medical Research Center (NMRC) Rockville, MD, 2003-2004

Research Associate

. Purified antibodies and antigens against Rickettsial Disease

through small column chromatography and HPLC utilizing affinity

chromatography, ion exchange, and gel filtration. Tested

antibody strength by ELISA to determine titer and purity of

antibody. Purified antibodies and antigens were provided to

other scientists towards a large scale determination of a

Rickettsial vaccine.

Johns Hopkins University School of Medicine Baltimore MD, 2001-2003

Graduate Research Assistant

. Regulated DNA transcription by controlling the acetylation of

histone proteins.

. Designed and synthesized a library of twelve compounds and

tested them as inhibitors of P300, a histone acetyl transferase

enzyme. Two were found to be potent inhibitors of P300. This

opportunity provided experience with solid phase organic

synthesis, peptide synthesis, and protein purification.

University of Maryland Baltimore County Baltimore MD, 2000-2001

Lab Assistant

. Worked independently to synthesize and discover two covalently

bound inhibitors for glyoxalase I, an enzyme of the citric acid

cycle.

. Utilized MALDI to confirm the affinity labels were covalently

bonded to glyoxalase I. Inhibitors of glyoxalse I have shown to

be selective anticancer drugs in tumor cell lines as amounts of

glyoxalase I in these cells are abnormally high.

University of Maryland at College Park, 1995-1997

Student Lab Assistant

. Completed a high school advanced science program in which I

worked with a local scientist 20 hours a week on an independent

research project.

. Documented results and prepared a dissertation of results from

an analysis of the effect of temperature on the spread and

growth of DC3000 strain of Psuedomonas syringae infection in

tomato and tobacco plants. My results were used to further

research on the genetic engineering on tobacco and tomato plants

to prevent crop loss.

TECHNICAL EXPERIENCE

. Structural Biology-Training in X-ray crystallography including growing

and optimizing crystals. Experience in structure refinement

. Protein Purification-Affinity chromatography, gel filtration, ion

exchange, experience troubleshooting the purification of difficult

protein

. Molecular Biology-Recombinant and traditional cloning, PCR,

sequencing, bacterial and yeast transformations, protein expression,

site-directed mutagenesis, yeast and bacterial cell cultures

. Immunology-Westerns, ELISA, antigen design, antibody and antigen

purification

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