Abstract

Continuance is requested of a historical, MERIT-designated award responsible for much of present-day understanding of the architecture and biogenesis of mycobacterial cell walls. The new thrust is the development of drug targets/screens to counteract multiple drug resistant TB and, tangentially, aid in definition of mechanisms of disease pathogenesis. The experimental approach centers on four major topics: 1. Knowledge of the pathways of linker unit (LU), arabinogalactan (AG) and lipoarabinomannan (LAM) biosynthesis warrants identification and production of the relevant glycosyltransferases through synthesis of sugar-donors, neoglycolipid-acceptors and photoaffinity probes and through genome database searches, and application of the results to cascade, acceptor-donor and single-enzyme high-throughput screens to assess compound, combinatorial and natural product libraries in conjunction with GlaxoWellcome. 2. The unique structures of the mycobacterial polyprenyl-P carriers and expected unique specificity of the isoprenol initiating reactions warrants analysis of these pathways in terms of the worth of such fundamental information and its possible application in drug development. 3. The extreme paucity of information on the peptidoglycan of M. tuberculosis invites an examination of basic structure and biosynthesis, assessment of conformation/ solution structure in the contexts of AG and cell wall proper by means of high resolution NMR, and definition of extra-membranous events such as polymerization, cross-linking and linkage to AG. 4. The recent recognition of the nature of the mycolic acid carrier, and the mycolyltransferase enzyme and the mechanism of Claisen condensation calls for an enzymatic and genetic approach to defining the mechanisms and essentiality of the terminal stages of mycolic acid synthesis, transport and deposition through recourse of drug-resistant genes to some novel mycolic acid antagonists, and """"""""knock-out"""""""" mutagenesis of these and other genes.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI018357-18
Application #
2469447
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
1981-01-01
Project End
2002-12-31
Budget Start
1998-01-01
Budget End
1998-12-31
Support Year
18
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Colorado State University-Fort Collins
Department
Microbiology/Immun/Virology
Type
Schools of Veterinary Medicine
DUNS #
112617480
City
Fort Collins
State
CO
Country
United States
Zip Code
80523

  Publications

Larrouy-Maumus, Gérald; Gilleron, Martine; Skovierová, Henrieta et al. (2015) A glycomic approach reveals a new mycobacterial polysaccharide. Glycobiology 25:1163-71
Ishizaki, Yoshimasa; Hayashi, Chigusa; Inoue, Kunio et al. (2013) Inhibition of the first step in synthesis of the mycobacterial cell wall core, catalyzed by the GlcNAc-1-phosphate transferase WecA, by the novel caprazamycin derivative CPZEN-45. J Biol Chem 288:30309-19
McShane, Helen; Jacobs, William R; Fine, Paul E et al. (2012) BCG: myths, realities, and the need for alternative vaccine strategies. Tuberculosis (Edinb) 92:283-8
Bhamidi, Suresh; Scherman, Michael S; Jones, Victoria et al. (2011) Detailed structural and quantitative analysis reveals the spatial organization of the cell walls of in vivo grown Mycobacterium leprae and in vitro grown Mycobacterium tuberculosis. J Biol Chem 286:23168-77
Skovierová, Henrieta; Larrouy-Maumus, Gérald; Pham, Ha et al. (2010) Biosynthetic origin of the galactosamine substituent of Arabinogalactan in Mycobacterium tuberculosis. J Biol Chem 285:41348-55
Amin, Anita G; Angala, Shiva K; Chatterjee, Delphi et al. (2009) Rapid screening of inhibitors of Mycobacterium tuberculosis growth using tetrazolium salts. Methods Mol Biol 465:187-201
Scherman, Hataichanok; Kaur, Devinder; Pham, Ha et al. (2009) Identification of a polyprenylphosphomannosyl synthase involved in the synthesis of mycobacterial mannosides. J Bacteriol 191:6769-72
Skovierová, Henrieta; Larrouy-Maumus, Gérald; Zhang, Jian et al. (2009) AftD, a novel essential arabinofuranosyltransferase from mycobacteria. Glycobiology 19:1235-47
Eoh, Hyungjin; Brennan, Patrick J; Crick, Dean C (2009) The Mycobacterium tuberculosis MEP (2C-methyl-d-erythritol 4-phosphate) pathway as a new drug target. Tuberculosis (Edinb) 89:1-11
Kaur, Devinder; Guerin, Marcelo E; Skovierová, Henrieta et al. (2009) Chapter 2: Biogenesis of the cell wall and other glycoconjugates of Mycobacterium tuberculosis. Adv Appl Microbiol 69:23-78

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