HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Köplin, R. Articles by Whitfield, C. Search for Related Content PubMed PubMed Citation Articles by Köplin, R. Articles by Whitfield, C. Social Bookmarking                      What's this?

Volume 272, Number 7, Issue of February 14, 1997 pp. 4121-4128
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

---

UDP-galactofuranose Precursor Required for Formation of the Lipopolysaccharide O Antigen of Klebsiella pneumoniae Serotype O1 Is Synthesized by the Product of the rfbD_KPO1 Gene

(Received for publication, September 16, 1996, and in revised form, November 11, 1996)

Reinhard Köplin , Jean-Robert Brisson ^§ and Chris Whitfield

From the Canadian Bacterial Diseases Network,  Department of Microbiology, University of Guelph, Guelph, Ontario, Canada, N1G 2W1, and the ^§ Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada, K1A 0R6

The O-side-chain polysaccharide in the lipopolysaccharide of Klebsiella pneumoniae O1 is based on a backbone structure of repeat units of [3)--D-Galf-(13)--D-Galp-(1]; this structure is termed D-galactan I. The rfb (O-antigen biosynthesis) gene cluster directs the synthesis of D-galactan I and consists of six genes termed rfbA-F_KPO1. In this paper we show that rfbD_KPO1 encodes a UDP-galactopyranose mutase (NAD(P)H-requiring) (EC 5.4.99.9), which forms uridine 5-(trihydrogen diphosphate) P--D-galactofuranosyl ester (UDP-Galf), the biosynthetic precursor of galactofuranosyl residues. The deduced amino acid sequence of rfbD_KPO1 shows 85% and 37.5% identity to the rfbD_KPO8 gene of K. pneumoniae serotype O8 and the glf gene of Escherichia coli, respectively. The molecular mass of the purified RfbD_KPO1 enzyme is 45 kDa as determined by SDS-polyacrylamide gel electrophoresis, while gel filtration revealed a molecular mass of 92 kDa, suggesting a dimeric structure for the native protein. The rfbD_KPO1 gene product interconverts uridine 5-(trihydrogen diphosphate) P--D-galactopyranosyl ester (UDP-Galp) and UDP-Galf. Unlike Glf, RfbD_KPO1 showed a requirement for NADH or NADPH, which could not be replaced by NAD or NADP. RfbD_KPO1 was used to synthesize milligram quantities of UDP-Galf, allowing this compound to be purified and fully characterized in an intact form for the first time. The structure of UDP-Galf was proven by NMR spectroscopy.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				P. S. Schmalhorst, S. Krappmann, W. Vervecken, M. Rohde, M. Muller, G. H. Braus, R. Contreras, A. Braun, H. Bakker, and F. H. Routier Contribution of Galactofuranose to the Virulence of the Opportunistic Pathogen Aspergillus fumigatus Eukaryot. Cell, August 1, 2008; 7(8): 1268 - 1277. [Abstract] [Full Text] [PDF]

				B. Kleczka, A.-C. Lamerz, G. van Zandbergen, A. Wenzel, R. Gerardy-Schahn, M. Wiese, and F. H. Routier Targeted Gene Deletion of Leishmania major UDP-galactopyranose Mutase Leads to Attenuated Virulence J. Biol. Chem., April 6, 2007; 282(14): 10498 - 10505. [Abstract] [Full Text] [PDF]

				T. Konishi, T. Takeda, Y. Miyazaki, M. Ohnishi-Kameyama, T. Hayashi, M. A. O'Neill, and T. Ishii A plant mutase that interconverts UDP-arabinofuranose and UDP-arabinopyranose Glycobiology, March 1, 2007; 17(3): 345 - 354. [Abstract] [Full Text] [PDF]

				S. M. Beverley, K. L. Owens, M. Showalter, C. L. Griffith, T. L. Doering, V. C. Jones, and M. R. McNeil Eukaryotic UDP-Galactopyranose Mutase (GLF Gene) in Microbial and Metazoal Pathogens Eukaryot. Cell, June 1, 2005; 4(6): 1147 - 1154. [Abstract] [Full Text] [PDF]

				E. F. Mulrooney, K. K. H. Poon, D. J. McNally, J.-R. Brisson, and J. S. Lam Biosynthesis of UDP-N-acetyl-L-fucosamine, a Precursor to the Biosynthesis of Lipopolysaccharide in Pseudomonas aeruginosa Serotype O11 J. Biol. Chem., May 20, 2005; 280(20): 19535 - 19542. [Abstract] [Full Text] [PDF]

				W. L. Miller, C. Q. Wenzel, C. Daniels, S. Larocque, J.-R. Brisson, and J. S. Lam Biochemical Characterization of WbpA, a UDP-N-acetyl-D-glucosamine 6-Dehydrogenase Involved in O-antigen Biosynthesis in Pseudomonas aeruginosa PAO1 J. Biol. Chem., September 3, 2004; 279(36): 37551 - 37558. [Abstract] [Full Text] [PDF]

				A. El Zoeiby, F. Sanschagrin, A. Darveau, J.-R. Brisson, and R. C. Levesque Identification of novel inhibitors of Pseudomonas aeruginosa MurC enzyme derived from phage-displayed peptide libraries J. Antimicrob. Chemother., March 1, 2003; 51(3): 531 - 543. [Abstract] [Full Text] [PDF]

				B. Kneidinger, K. O'Riordan, J. Li, J.-R. Brisson, J. C. Lee, and J. S. Lam Three Highly Conserved Proteins Catalyze the Conversion of UDP-N-acetyl-D-glucosamine to Precursors for the Biosynthesis of O Antigen in Pseudomonas aeruginosa O11 and Capsule in Staphylococcus aureus Type 5. IMPLICATIONS FOR THE UDP-N-ACETYL-L-FUCOSAMINE BIOSYNTHETIC PATHWAY J. Biol. Chem., January 31, 2003; 278(6): 3615 - 3627. [Abstract] [Full Text] [PDF]

				C. Schaffer, T. Wugeditsch, P. Messner, and C. Whitfield Functional Expression of Enterobacterial O-Polysaccharide Biosynthesis Enzymes in Bacillus subtilis Appl. Envir. Microbiol., October 1, 2002; 68(10): 4722 - 4730. [Abstract] [Full Text] [PDF]

				L. E. Hancock and M. S. Gilmore The capsular polysaccharide of Enterococcus faecalis and its relationship to other polysaccharides in the cell wall PNAS, February 5, 2002; 99(3): 1574 - 1579. [Abstract] [Full Text] [PDF]

				D. C. Crick, S. Mahapatra, and P. J. Brennan Biosynthesis of the arabinogalactan-peptidoglycan complex of Mycobacterium tuberculosis Glycobiology, September 1, 2001; 11(9): 107R - 118R. [Abstract] [Full Text] [PDF]

				F. Pan, M. Jackson, Y. Ma, and M. McNeil Cell Wall Core Galactofuran Synthesis Is Essential for Growth of Mycobacteria J. Bacteriol., July 1, 2001; 183(13): 3991 - 3998. [Abstract] [Full Text] [PDF]

				S. Guan, A. J. Clarke, and C. Whitfield Functional Analysis of the Galactosyltransferases Required for Biosynthesis of D-Galactan I, a Component of the Lipopolysaccharide O1 Antigen of Klebsiella pneumoniae J. Bacteriol., June 1, 2001; 183(11): 3318 - 3327. [Abstract] [Full Text]

				M. Graninger, B. Nidetzky, D. E. Heinrichs, C. Whitfield, and P. Messner Characterization of dTDP-4-dehydrorhamnose 3,5-Epimerase and dTDP-4-dehydrorhamnose Reductase, Required for dTDP-L-rhamnose Biosynthesis in Salmonella enterica Serovar Typhimurium LT2 J. Biol. Chem., August 27, 1999; 274(35): 25069 - 25077. [Abstract] [Full Text] [PDF]

				P. Chen and W. R. Bishai Novel Selection for Isoniazid (INH) Resistance Genes Supports a Role for NAD+-Binding Proteins in Mycobacterial INH Resistance Infect. Immun., November 1, 1998; 66(11): 5099 - 5106. [Abstract] [Full Text] [PDF]

				K. Mikusova, T. Yagi, R. Stern, M. R. McNeil, G. S. Besra, D. C. Crick, and P. J. Brennan Biosynthesis of the Galactan Component of the Mycobacterial Cell Wall J. Biol. Chem., October 20, 2000; 275(43): 33890 - 33897. [Abstract] [Full Text] [PDF]