Elsa D. Garcin, Ph.D.

The Scripps Research Institute

Department of Molecular Biology, MB4

10550 North Torrey Pines, La Jolla, CA

elsag@scripps.edu

Personal     
Date of Birth:   July 22, 1971
Citizenship:      France

Summary

Research associate with a PhD and 10 years experience in protein crystallography. Particular expertise in structure-based drug design. Dedicative and innovative team player with a comprehensive background in protein biochemistry and spectroscopic methods.

 

·       PhD in Physics with a strong research background in biochemistry, x-ray crystallography, structure-based drug design, and spectroscopic methods.

·       Obtained a tenure-track faculty position as Assistant Professor in Biochemistry in the Department of Chemistry and Biochemistry at University of Maryland Baltimore County.

·       Developing new projects aiming to elucidate nitrogen oxide modifications of protein structure and protein-protein interactions in cardiovascular health and disease.  These projects focus on key protein targets with important therapeutic implications in cardiovascular diseases, including heart failure, diabetes, metabolic disease, and neurodegenerative vascular disorders.

·       Extensive experience in molecular biology, protein expression and purification, crystallography, anaerobic techniques, spectroscopic and biochemical characterizations, computational and Small-Angle X-ray Scattering techniques, and Deuterium/Hydrogen Exchange Mass Spectrometry.

·       Excellent written and oral communication skills, strong collaborative skills, extensive teaching experience with undergraduate students, supervised and mentored high school, undergraduate, and graduate students, and research technicians, trained postdoctoral researchers.

·                     Designed and wrote competitive funded fellowship (AHA), funded NIH R01 grant (under E. Getzoff)
 

Experience

University of Maryland, Baltimore County
Department of Chemistry and Biochemistry
 
Assistant professor                                                  08/2008-
 
The Scripps Research Institute, 
Department of Molecular Biology, La Jolla, CA
 
Senior Postdoctoral Fellow                                           07/2007-07/2008
(The Skaggs Institute for Chemical Biology and NIH)  
        Laboratories of Profs. John Tainer and Elizabeth Getzoff
 
Research Associate                                                   2005 – 2007
(The Skaggs Institute for Chemical Biology and NIH)
        Laboratories of Profs. John Tainer and Elizabeth Getzoff
 
Research Associate                                                   2004 – 2005
(NIH)
        Laboratory of Dr. Elizabeth Getzoff
 
American Heart Association Postdoctoral Research Fellow              2002 – 2004
 
Research Associate                                                   1999 – 2002
(NIH)
        Laboratory of Dr. Elizabeth Getzoff

 

·           Performed structural, mutagenesis and kinetics studies for the three-zinc endonuclease IV to address mechanisms for cleavage and coordination of product handoff in the base-excision repair pathway for DNA repair.

·           Determined >30 crystal structures of nitric oxide synthase inhibitor complexes in collaboration with AstraZeneca (UK) and MCR Inc. (Toronto, Canada).

·           Elucidated the first structure of the intact nitric oxide synthase electron-transferring reductase module, and Identified new regulatory elements for electron transfer regulation.

·           Successfully initiated and completed project to investigate redox states of the nitric oxide synthase reductase module generated in synchrotron x-ray beam and monitored with UV-visible absorbance microspectrophotometry.

·           Designed, cloned, and purified new mutants in full-length NOS relevant in regulation and assembly.

·           Initiated Deuterium/Hydrogen Exchange (DXMS) mass spectrometry studies on NOS in collaboration with Dr Virgil Woods, at UCSD.

·           Designed and executed small-angle x-ray scattering (SAXS) experiments to investigate holo-NOS conformational changes upon calmodulin binding.

·           Supervised and trained graduate students and postdoctoral researchers in the use of the laboratory microspectrophotometer.

·           Was awarded 1 competitive postdoctoral fellowship and ranked first among all applicants.

·           Wrote as part of team 1 funded NIH RO1 grant.

·           Designed and was primary author for funded NIH RO1 grant (under E. Getzoff, submitted March 2006, awarded a 130 score and 6.0 percentile).

·           Directed, trained, and supervised undergraduate, high school, and rotating graduate students and supervised technician.

·           Wrote proposals for and was awarded protein crystallography and small-angle x-ray scattering beamtime at national synchrotron facilities (SSRL, ALS, APS, and ESRF).

 

The Structural Biology Institute Jean-Pierre Ebel, Grenoble, France
 
Postdoctoral Fellow                                                  1998 - 1999
Department of Molecular Biology
        Laboratory of Dr Giuseppe Zaccai
 
“Contrat de Formation par la Recherche” (CEA) Graduate Fellow        1994 - 1998
Department of Protein Crystallogenesis and Crystallography
        Laboratory of Dr. Juan-Carlos Fontecilla-Camps
 
Junior Lecturer (biochemistry and Spectroscopic Methods)             1997 - 1998
Joseph Fourier University, France

 

·           Successfully initiated and completed project using anaerobic glove-box and x-ray crystallography to investigate redox states of [NiFeSe] hydrogenase from Desulfomicrobium baculatum.

·           Identified bimetallic nature of active site of [NiFe] hydrogenase from Desulfovibrio gigas by anomalous dispersion.

·           Successfully produced and collected x-ray, neutron, and small-angle x-ray scattering data for malate dehydrogenase from extreme halophile Haloharcula marismortui

·           Instructed undergraduates in biochemistry, Sr. level spectroscopic methods, and Sr. level biochemistry laboratory classes.

·           Directed, trained, and supervised undergraduate student

·            

Education

·  Ph.D., Physics/Biochemistry                                                                                                                     June 1998

“Cryocrystallographic Studies of the active site of the Nickel-iron hydrogenases from Desulfovibrio gigas and Desulfomicrobium baculatum”

Joseph Fourier University (France), Summa Cum Laude

Mentor: Juan-Carlos Fontecilla-Camps, PhD.

·  Master’s 2 degree in Biological Crystallography and NMR                                                                          June 1994

Joseph Fourier University (France) Magna Cum Laude (ranked 1st)

·  Master’s 1 degree in Biochemistry                                                                                                             June 1993

University of Marseille (France), Summa Cum Laude (ranked 1st)

·  B. Sc. Biochemistry                                                                                                                                 June 1992

University of Marseille (France) Magna Cum Laude

 

Research and Professional Experience

 
2007           Senior Postdoctoral Fellow at the Scripps Research Institute
1999-2007      Research Associate at the Scripps Research Institute
1998           6 months training course in purification, neutrons and small angle scattering techniques (LBM-IBS) 
1994-1998      Thesis work in Crystallography (LCCP-IBS), financed by CEA-Grenoble 
1994           5 months training course in Protein Crystallography (LCCP-IBS, Grenoble, FR) 
1993           4 weeks training course in EPR (University of Marseille, FR) 




 

Skills:                                                                                              

·       Biochemistry: Extensive experience with protein expression (E. coli), purification (FPLC, affinity and size-exclusion chromatography), characterization (SDS and native PAGE, Western Blot, UV-visible spectroscopy, light scattering). Extensive experience with crystal microspectrophotometer. Expert in anaerobic manipulation of proteins and protein crystals. Experience with DXMS sample preparation.

·       Peer Review: Participated in peer review of journal article submissions including J. Biol. Chem, Biochemistry, and J. Med. Chem. Peer reviewed pre-submission of revised edition of book for Elsevier. Participated in peer review of National Science Foundation grant applications.

·       Other: Experience with programs for electrostatic potential calculations (UHBD), SAXS and DXMS data processing. Strong analytical abilities, communication and organizational skills.

 

Teaching Experience: 

The Scripps Research Institute
·                     Supervisor for rotating graduate student                                                                                     2007
·                     Supervised and trained graduate students and postdoctoral fellows in the use 
               of the laboratory microspectrophotometer, anaerobic glove-box and cryocrystallography tools              2007
·                     Supervisor for research technician                                                                             2006-present
·                     Supervisor for high-school summer student                                                                                2005
·                     Supervisor for research technician                                                                             1999-2001
 
Joseph Fourier University, Grenoble (France)
·                     Junior lecturer in Enzymology laboratory classes for Sr. undergraduate students                 1998
“Proteins: purification, enzymology, kinetics” - 54 hours.
Biochemical characterization of catalase: how to design, analyze, and interpret experiments
·                     Junior lecturer in Spectroscopic Methods for Sr. undergraduate students                                           1997
UV and IR methods- 24 hours            
·                     Junior Lecturer in Enzymology laboratory class for Sr. undergraduate students                                  1997
“Proteins: purification, enzymology, kinetics”- 27 hours.
 
The Structural Biology Institute Jean-Pierre Ebel, Grenoble (France)
·                     Trainee supervisor for an undergraduate student (6 months)                                                            1995-1996

 

 

Awards

2008                             Member of the American Heart Association

2007-2008                      Member of the Society for Free Radical in Biology and Medicine         

2006-2007                      Skaggs Training Grant                 

2005                             Travel Award from the 15th International Symposium on Flavins and Flavoproteins

2003                             Lievre Family Special Award for being top-ranked postdoc of the American Heart Association-Western States Affiliate

2002                             Travel Award from US National Committee for Crystallography and NASA

2002-2004                      PostDoctoral Fellowship of the American Heart Association-Western States Affiliate

1995-1998                      Award of a Contrat de Formation par la recherche (CEA- Grenoble)

 

Areas of interest

I am a talented crystallographer, with a strong background in biochemistry and spectroscopic methods. My 8 years of postdoctoral studies in protein X-ray crystallography have focused on Nitric Oxide Synthase structural studies in the laboratory of Prof. Elizabeth Getzoff, at the Scripps Research Institute. I am currently engaged in a project involving several aspects of the NOS studies that include structure-based design of isozyme-specific inhibitors, structural studies of the electron donor reductase module, and assembly and regulation of the holo-enzyme, by coupling molecular biology, high-resolution protein crystallography, spectroscopic, biochemical, computational and Small-Angle X-ray Scattering (SAXS) techniques, and Deuterium/Hydrogen Exchange Mass Spectrometry (DXMS).

Publications

 

Garcin E.D., A.S. Arvai, R.J. Rosenfeld, M.D. Kroeger, B.R. Crane, G. Andersson, G. Andrews, P.J. Hamley, P.R. Mallinder, D.J. Nicholls, S.A. St-Gallay, A.C. Tinker, N.P. Gensmantel, A. Mete, D.R. Cheshire, S. Connolly, D.J. Stuehr, A. Åberg, A.V. Wallace, J.A. Tainer, E.D. Getzoff. Anchored plasticity opens doors for selective inhibitor design in NOS. (Submitted to Nature, pending revisions).

 

Garcin E.D., Hosfield D.J., Haas B.J., Björas M., Cunningham R., Tainer J.A. Endonuclease IV catalytic mechanism and coordination of DNA base excision repair from enzyme:DNA crystal structures, mutational analyses, and kinetics (Submitted to Nat. Struct. Mol. Biol., pending revisions).

Rameau GA, Tukey DS, Garcin-Hosfield ED, Titcombe RF, Misra C, Kathri L, Getzoff ED, Ziff E.B. "Biphasic coupling of neuronal nitric oxide synthase phosphorylation to the NMDA receptor regulates AMPA receptor trafficking and neuronal cell death." Journal of Neuroscience 27 (13), (2007), 3445-3455

Panda K, Haque MM, Garcin-Hosfield ED, Durra D, Getzoff ED, Stuehr DJ. "Surface charge interactions of the FMN module govern catalysis by nitric-oxide synthase." J. Biol. Chem. 281 (48), (2006), 36819-36827

Tiso M, Konas DW, Panda K, Garcin ED, Sharma M, Getzoff ED, Stuehr DJ. "C-terminal tail residue ARG1400 enables NADPH to regulate electron transfer in neuronal nitric oxide synthase." J. Biol. Chem. 280 (47), (2005), 39208-39219

Volbeda, A., Martin, L., Cavazza, C., Matho, M., Faber, B.W., Roseboom, W., Albracht, S.P.J., Garcin, E., Rousset, M., Fontecilla-Camps, J.-C. "Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases". Journal of Biological Inorganic Chemistry 10 (3), (2005), 239-249

Wei, C.-C., Wang, Z.-Q., Durra, D., Hemann, C., Hille, R., Garcin, E.D., Getzoff, E.D., Stuehr, D.J. "The three nitric-oxide synthases differ in their kinetics of tetrahydrobiopterin radical formation, heme-dioxy reduction, and arginine hydroxylation". J. Biol. Chem. 280 (10), (2005), 8929-8935

Garcin E.D., Bruns C.M., Lloyd S.J., Hosfield D.J., Tiso M., Gachhui R., Stuehr D.J., Tainer J.A. and Getzoff E.D. Structural basis for isozyme-specific regulation of Electron Transfer in Nitric Oxide Synthase". J. Biol. Chem. 279 (3), (2004), 37918-37927

Rosenfeld R.J., Garcin E.D.,Panda K., Andersson G., Aberg A., Wallace A.V., Morris G.M., Olson A.J., Stuehr D.J., Tainer J.A., Getzoff E.D., "Conformational Changes in Nitric Oxide Synthases Induced by Chlorzoxazone and Nitroindazoles: Crystallographic and Computational Analyses of Inhibitor Potency" Biochemistry, 41(47), (2002), 13915-13925

Richard S.B., Madern D., Garcin E., Zaccai G., Halophilic adaptation: Novel solvent protein interactions observed in the 2.9 and the 2.6 Angstrom resolution structures of the wild type and and a mutant of Malate Dehydrogenase from Haloarcula Marismortui" Biochemistry, 39 (5), (2000), 992-1000.

E. Garcin, X. Vernede, C. Hatchikian, A. Volbeda, M. Frey, J.-C. Fontecilla-Camps, "The Crystal Structure of a reduced [NiFeSe] hydrogenase provides an image of the activated catalytic center" Structure, 7 (5), (1999), 557-566.

Garcin E., Montet Y., Volbeda A., Hatchikian E.C., Frey M., Fontecilla-Camps J.-C., "Structural Bases for the catalytic mechanism of [NiFe] hydrogenases", Biochem. Soc. Trans., 26 (3), (1998), 396-401.

Y. Montet, E. Garcin, A. Volbeda, C. Hatchikian, M. Frey, J.-C. Fontecilla-Camps, "Structural bases for the catalytic mechanism of NiFe hydrogenase", Pure & Appl. Chem., 70, 1 (1998), 25-31.

J.-C. Fontecilla-Camps, M. Frey, E. Garcin, C. Hatchikian, Y. Montet, C. Piras, X. Vernede, A. Volbeda, "Hydrogenase: a hydrogen-metabolizing enzyme. What do the crystal structures tell us about its mode of action ?", Biochimie, 79 (1997), 661-666.

Volbeda A. Garcin E., Piras C., de Lacey A. I., Fernandez V. M., Hatchikian E. C., Frey M., Fontecilla-Camps J.-C., "Structure of the [NiFe] hydrogenase active site: evidence for biologically uncommon Fe ligands", J. Am. Chem. Soc., 118,51 (1996), 12989-12996.

Meetings and Workshop

2007, Oct           The Society of Fellows Fall Symposium, TSRI, La Jolla

Poster:  “Overcoming active-site conservation to create isozyme specificity: Inhibitors, structures and mutants of nitric oxide synthase”.

2007, Feb          Nitric Oxide GRC (Ventura, CA)
Poster: Structural Insights into the conformational selectivity of highly specific inducible nitric oxide synthase inhibitors

2006, June         Fourth International Conference on Biochemistry and Therapeutic Applications of Nitric oxide (Monterey, CA)

 Hot Topics: Structural insights into NOS regulatory mechanisms:  Molecular shape changes in NOS upon calmodulin binding revealed by small-angle x-ray scattering (SAXS)

2005, April        15th International Symposium on Flavins and Flavoproteins (Shonan Village, Japan)
Poster: Structural Basis for isozyme-specific regulation of electron transfer in Nitric oxide Synthase

2005, March       WCPW (Asilomar Conference Center, Pacific Grove)
Invited Talk: Structural Basis for isozyme-specific regulation of electron transfer in Nitric oxide Synthase

2004, June         "GTBIO Lyon" (France)
Invited talk: Structural studies of Nitric Oxide Synthases: Structure-Based Drug design and isozyme-specific regulation of electron transfer

2003, June         "Nitric Oxide: A Janus molecule" (Roma, Italy)

Invited talk: Crystal Structures of Nitric Oxide Synthase oxygenase and reductase modules: Implications for holo-enzyme Assembly and Regulation

2003, May         "The Society of Fellows Spring Symposium" (TSRI, La Jolla)
Poster: Crystal Structures of Nitric Oxide Synthase oxygenase and reductase modules: Implications for holo-enzyme Assembly and Regulation

2003, March       "The 16th West Coast Protein Crystallography Workshop" (Asilomar Conference Center, Pacific Grove, CA)

Poster: Crystal Structures of Nitric Oxide Synthase oxygenase and reductase modules: Enzyme Mechanism and Regulation of Electron Transfer

2003, Feb          "Gordon Conference: Nitric Oxide" (Ventura, CA)

Poster: Crystal Structures of Nitric Oxide Synthase oxygenase and reductase modules: Enzyme Mechanism and Regulation of Electron Transfer

2002, Sep          "Young Investigator Forum-American Heart Association (San Diego, CA)

Invited Talk: Crystal Structures of Nitric Oxide Synthase: Structure Based Drug Design and Regulation of Electron Transfer

2002, Aug          "XIXth IUCR Meeting, (Geneva, Switzerland)

Poster: Crystal Structures of Nitric Oxide Synthase oxygenase and reductase modules: Enzyme Mechanism and Regulation of Electron Transfer

1999, Feb          "ESRF Users' Meeting" (Grenoble, France)
Invited talk: Determination of metallic sites in proteins by anomalous dispersion experiment

1998, Feb          "Meeting of the French Crystallographic Association AFC 98" (Orleans, France)

Poster: Structure de l'hydrogenase a [NiFeSe] de Desulfomicrobium baculatum

1998, Jan           Meeting of the french group "Physique et Chimie des metalloproteines et de leurs analogues de synthese" (Valloire, France)

Invited talk: Structure cristallographique de l'hydrogenase a [NiFeSe] de Desulfomicrobium baculatum

1997, Oct           "DBMS/IBS Meeting" (Grenoble, France)
Invited talk: Structure cristallographique a 2.2 Angstroms de resolution de l'hydrogenase a [NiFeSe] de Desulfomicrobium baculatum

1997, Aug          "European Crystallographic meeting" (Lisboa, Portugal)
Poster: Structural studies of the [NiFe] hydrogenase from Desulfovibrio gigas

1997, Jul           European Commission and COST Conference, "Vth International Conference on the Molecular Biology of hydrogenases" (Albertville, France)

Invited talk: Crystallographic studies of the hydrogenase from Desulfomicrobium baculatum

1997, Jan           "Meeting of the GDR-CNRS Metalloproteines et analogues" (Toulouse, France)
Poster: Etudes structurales du site actif de l'hydrogenase de Desulfovibrio gigas

1996, Sep          "Meeting of the French Crystallographic Association AFC 96" (Toulouse, France)
Invited talk: Etudes cryocristallographiques du site actif de l'hydrogenase de Desulfovibrio gigas

1996, May         "48 hours of Structural Biology" (Autrans, France)
Invited talk: Determination de la nature du site catalytique bimetallique par dispersion anomale de l'hydrogenase de Desulfovibrio gigas

1996, May         COST Workshop "Hydrogenase structures and functional implications" (Grenoble, France)
Invited talk: Cryocrystallographic studies of the redox states of the active site of D. gigas hydrogenase

1996, Jan           "Macromolecular refinement" (Chester, UK)

1995, Aug          "Biophysics and Synchrotron Radiation"(Grenoble, France)

1995, May         Biotech meeting "Biomimetic chemistry of hydrogenases"(Madrid, Spain)

1994, Oct           Biotech meeting "Design of improved biomimetic catalysts through structural and functional studies on nickel-containing hydrogenases"(Grenoble, France)

Crystal Structures determined and deposited as Protein Data Bank Entries

1M8D (2002) murine iNOSox, Chlorzoxazone, 2.35 Å Resolution 
1M8E (2002) murine iNOSox, 7-Nitroindazole, 2.9Å Resolution 
1M8H (2002), murine iNOSox, 6-Nitroindazole, 2.85Å Resolution 
1M8I (2002) murine iNOSox, 5-Nitroindazole, 2.8Å Resolution 
1M9T (2002) murine iNOSox, 3-Bromo,7-Nitroindazole, 2.4 Å Resolution 
1M9J (2002) human eNOSox, Chlorzoxazone, 2.4Å Resolution 
1M9K (2002) human eNOSox, 7-Nitroindazole, 2.01Å Resolution 
1M9M (2002) human eNOSox, 6-Nitroindazole, 1.96Å Resolution 
1M9Q (2002) human eNOSox, 5-Nitroindazole, 2.01Å Resolution 
1M9R (2002) human eNOSox, 3-Bromo,7-Nitroindazole, 2.56Å Resolution 
1TLL (2004) rat nNOSred 2.3Å Resolution
2A44 (2005) human iNOSox, AR-C95791, 2.1Å resolution 
2A4P (2005) murine iNOSox, AR-C102222, 2.6Å resolution
2A4S (2005) bovine eNOSox, AR-C95791, 2.5Å resolution
2A4U (2005) murine iNOSox, AR-C132283, 2.3Å resolution
2A5O (2005) murine iNOSox, AR-C118901, 2.5Å resolution
2A5Q (2005) murine iNOSox, AR-C124355, 2.6Å resolution
2F6O (2005) murine iNOSox, AR-C130232, 2.2Å resolution
2F85 (2005) murine iNOSox, AR-C95791, 2.0Å resolution
2FAN (2005) murine iNOSox, AR-C125813, 2.2Å resolution
2FEV (2005) murine iNOSox, AR-C94864, 2.9Å resolution
2F7H (2005) murine iNOSox, AR-C120011, 2.05Å resolution
2F7U (2005) murine iNOSox, 4MAP, 2.7Å resolution
2NQ9 (2006) Escherichia coli endonuclease IV Y72A mutant bound to damaged DNA, 1.45 Å resolution
2NQH (2006) E. coli endonuclease IV E261Q mutantbound to phosphate, 1.1 Å resolution
2NQJ (2006) E. coli endonuclease IV E261Q mutant bound to damaged DNA, 2.45 Å resolution

 

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e-mail:elsag@scripps.edu