Abstract

The specific aims in this grant are directed towards understanding the structure and mechanism of an essential bacterial glycosyltransferase that is involved in the biosynthesis of peptidoglycan. This enzyme, MurG, is found in all organisms that synthesize peptidoglycan and is a target for the design of new antibiotics. We developed the first direct assay to study MurG and recently solved the crystal structure of E. coli MurG. This first MurG structure, combined with sequence data on other glycosyltransferases, has revealed that MurG is a paradigm for a large family of metal ion-independent glycosyltransferases found in both eukaryotes and prokaryotes. Therefore, the studies proposed here are not only relevant to understanding an important antibacterial target, but will shed light on the structure and mechanism of an entire class of glycosyltransferases. Our long term goals are to learn how to design inhibitors for this family of glycosyltransferases and to learn how to manipulate substrate specificity. The former goal could lead to the development of new drugs; the latter should facilitate efforts to understand the cellular roles of glycosyltransferases and their products.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
7R01AI044854-07
Application #
6938428
Study Section
Special Emphasis Panel (ZRG1-SSS-B (01))
Program Officer
Peters, Kent
Project Start
1999-03-01
Project End
2007-02-28
Budget Start
2004-07-01
Budget End
2005-02-28
Support Year
7
Fiscal Year
2004
Total Cost
$56,655
Indirect Cost

  Institution

Name
Harvard University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Leimkuhler, Catherine; Fridman, Micha; Lupoli, Tania et al. (2007) Characterization of rhodosaminyl transfer by the AknS/AknT glycosylation complex and its use in reconstituting the biosynthetic pathway of aclacinomycin A. J Am Chem Soc 129:10546-50
Love, Kerry Routenberg; Swoboda, Jonathan G; Noren, Christopher J et al. (2006) Enabling glycosyltransferase evolution: a facile substrate-attachment strategy for phage-display enzyme evolution. Chembiochem 7:753-6
Zhang, Yu-Hui; Ginsberg, Cynthia; Yuan, Yanqiu et al. (2006) Acceptor substrate selectivity and kinetic mechanism of Bacillus subtilis TagA. Biochemistry 45:10895-904
Yuan, Yanqiu; Chung, Hak Suk; Leimkuhler, Catherine et al. (2005) In vitro reconstitution of EryCIII activity for the preparation of unnatural macrolides. J Am Chem Soc 127:14128-9
Gross, Benjamin J; Kraybill, Brian C; Walker, Suzanne (2005) Discovery of O-GlcNAc transferase inhibitors. J Am Chem Soc 127:14588-9
Lee, Ho Young; Chung, Hak Suk; Hang, Chao et al. (2004) Reconstitution and characterization of a new desosaminyl transferase, EryCIII, from the erythromycin biosynthetic pathway. J Am Chem Soc 126:9924-5
Helm, Jeremiah S; Hu, Yanan; Chen, Lan et al. (2003) Identification of active-site inhibitors of MurG using a generalizable, high-throughput glycosyltransferase screen. J Am Chem Soc 125:11168-9
Chen, Lan; Men, Hongbin; Ha, Sha et al. (2002) Intrinsic lipid preferences and kinetic mechanism of Escherichia coli MurG. Biochemistry 41:6824-33
Ye, X Y; Lo, M C; Brunner, L et al. (2001) Better substrates for bacterial transglycosylases. J Am Chem Soc 123:3155-6
Fan, E; Merritt, E A; Verlinde, C L et al. (2000) AB(5) toxins: structures and inhibitor design. Curr Opin Struct Biol 10:680-6