Celia W. GouldingAssistant Professor, Molecular Biology and Biochemistry Assistant Professor, Pharmaceutical Sciences |
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Research Interests |
structural biology, biochemistry, proteomics, microbiology, X-ray crystallography | |
| URL | LabWebsite | |
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Research Abstract |
The principal focus of my lab is to utilize proteomic and crystallographic techniques to elucidate and characterize new systems of protein complexes in Mycobacterium tuberculosis. Our overall goal is to create a systems approach to shift the focus of structural biology from a single protein to molecular assemblies. The systems of immediate interest contain potential anti-TB protein drug targets and protein membrane components. Novel Mycobacterial Heme Uptake System Iron is an essential metal for life. Mycobacteria must import iron from its host. Molecules involved in iron chelating pathways are well characterized, such as those involving exochelins and siderophores. Recently, it has been shown that during the early stages of S. aureus infection the major source of nutrient iron is heme rather than transferrin iron. A potential mycobacterial secreted hemophore (heme scavenging protein) has been identified in mycobacteria. Also, a potential cytosolic heme-degrading protein has been identified. Hence, a novel mcobacterial heme uptake system may exisit. My laboratory will be dissecting this pathway both structrually and biochemically. Thus far, the X-ray crystal structure of the potential hemophore has been deteremined, and a potential heme uptake membrane protein has been identified by mass spectrometry. Investigation into this uptake system is on-going. A System Involved in Bacterial-Host Cell Interactions. In the genome organization, surrounding the Cfp10/Esat6 genes, there is a conserved combination of genes which include secreted serine proteases, putative ABC-transporters, ATP-binding proteins and other membrane-associated proteins. These membrane-associated and energy-providing proteins may function to secrete the members of the Cfp10/Esat6 protein families, and the proteases may be involved in processing the secreted peptide. This cluster of genes encodes a potential new secretion machine where one could infer that these gene products form and act as a protein complex. My laboratory intends to express these membrane proteins and characterize their interaction partners and structures to shed light on this novel secretion system. Further studies will include the investigation into protein-protein interactions of Esat6/Cfp10 proteins in complex and alone with host proteins, which will be carried out in human macrophage cell line. Disulfide Bond Isomerase System Disulfide bond-forming (Dsb) proteins have been shown to be involved in virulence in many pathogenic bacteria. They are oxidoreductase proteins that have a variety of functions including chaperone activity, electron transfer and disulfide bond isomerase activity. It has been predicted that of the 161 potential secreted proteins of M. tuberculosis approximately 60 % of these contain at least one disulfide bon. Hence, Dsb proteins which assist in folding secreted proteins into their correct conformation and assist in disulfide bond formation are of great importance for the survival of M. tuberculosis. Utilizing bioinformatics, two secreted Dsb proteins have been identified in M. tuberculosis, one of which is my target protein to investigate the secreted disulfide bond isomerase system of mycobacteria. The secreted proteins which these two Dsb proteins interact with is presently under investigation. |
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| Publications | N. Chim, L.M. McMath, M. Beeby and C.W. Goulding, “Advances in Mycobacterium tuberculosis structural genomics: investigating potential chinks inthe armor of a deadly pathogen.” 2009, Infectious Disorders – Drug Targets, Oct1 epub. | |
| I. H. Khan, R. Raindran, J. Yee, M. Ziman, D.M. Lewisohn, M.L. Gennaro, J.L. Fylnn, C.W. Goulding, K. Deriemer, N.W. Lerche and P.A. Luciw,, “Profiling Antibodies to Mycobacterium tuberculosis (M. tb.) by Multiplex Microbead Suspension Arrays for Serodiagnosis of TB.”, 2008, Clin Vaccine Immunol, 15, 433-8. | ||
| B.G. Fox, C Goulding, M.G. Malkowski, L. Stewart and A. Deacon. “Structural genomics: from genes to structures with valuable materials and many questions in between”. 2008, Nature Methods, 52: 129-32. | ||
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“The structure and computational analysis of Mycobacterium tuberculosis protein CitE suggest a novel enzymatic function” Celia W. Goulding, Peter M. Bowers, Brent Segelke, Tim Lekin, Chang-Yub Kim, Thomas C. Terwilliger and David Eisenberg, J. Mol. Biol, 2007 Jan 12;365 :275-83 "A Novel Inhibitor of Mycobacterium tuberculosis Pantothenate Synthetase." White EL, Southworth K, Ross L, Cooley S, Gill RB, Sosa MI, Manouvakhova A,Rasmussen L, Goulding C, Eisenberg D, Fletcher TM 3rd. J Biomol Screen. 2006 Dec 14; [Epub] “Structural proteomics and computational analysis of a deadly pathogen: combating Mycobacterium tuberculosis from multiple fronts”, Michael Strong and Celia W Goulding, Methods Biochem Anal. 2006; 49:245-69. “Regulation by Oligomerization in a Mycobacterial Folate Biosynthetic Enzyme“, Celia W. Goulding, Marcin I. Apostol, Angineh Parseghian, Martin Philips and David Eisenberg, 2005, J. Mol. Biol. 349, 61-72. "Gram-positive DsbE proteins function differently to gram-negative DsbE homologs: A structural and functional analysis of secreted Mycobacterium tuberculosis DsbE", Celia W. Goulding, Marcin I. Apostol, Angineh Parseghian, Marila Gennaro and David Eisenberg J. Biol. Chem, 2004, 279, 3516-3524. “Protein Production in E. coli for Structural Studies”, Celia W. Goulding and L. Jeanne Perry J. of Structural Biology, 2003,142, 133-143. “The primary mechanism of attenuation of bacillus Calmette-Guerin is a loss of secreted lytic function required for invasion of lung interstitial tissue”, Tsungda Hsu, Suzanne M. Hingley-Wilson, Bing Chen, Paul Morin, Carolyn B. Marks, Celia Goulding, Mari Gingery, David Eisenberg, Robert G. Russell, Steven C. Derrick, Frank M. Collins, Sheldon L. Morris, C. Harold King and William R. Jacobs, Jr, Proc Natl Acad. Sci., 2003, 100, 12420-12425. "Crystal Structure of MPT63 – one of the three major secreted protein of Mycobacterium tuberculosis", Celia W. Goulding, Angineh Parseghian, Michael Sawaya, Duilio Cascio, Marcin I. Apostol, Marila Gennaro and David Eisenberg, Protein Science, 2002, 11, 2887-93. “Thiol:disulfide exchange in an immungloulin-like fold: Structure of the N-terminal domain of DsbD”, Celia W. Goulding, Michael Sawaya, Angineh Parseghian, Vincent Lim, David Eisenberg and Dominique Missiakas, Biochemistry, 2002 41, 6920-7. |
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Professional Society |
ACA |
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| Graduate Programs |
Structural Biology and Molecular Biophysics Cellular and Molecular Biosciences Medicinal Chemistry and Pharmacology |
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| Link to this profile | http://www.faculty.uci.edu/profile.cfm?faculty_id=5384 | |
| Last updated | 10/17/2009 | |