|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 281, Issue 2, 723-732, January 13, 2006
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Functional Characterization of Dehydratase/Aminotransferase Pairs from Helicobacter and Campylobacter
ENZYMES DISTINGUISHING THE PSEUDAMINIC ACID AND BACILLOSAMINE BIOSYNTHETIC PATHWAYS*
From the Institute for Biological Sciences, National Research Council, Ottawa, Ontario K1A OR6, Canada
Helicobacter pylori and Campylobacter jejuni have been shown to modify their flagellins with pseudaminic acid (Pse), via O-linkage, while C. jejuni also possesses a general protein glycosylation pathway (Pgl) responsible for the N-linked modification of at least 30 proteins with a heptasaccharide containing 2,4-diacetamido-2,4,6-trideoxy-
-D-glucopyranose, a derivative of bacillosamine. To further define the Pse and bacillosamine biosynthetic pathways, we have undertaken functional characterization of UDP--D-GlcNAc modifying dehydratase/aminotransferase pairs, in particular the H. pylori and C. jejuni flagellar pairs HP0840/HP0366 and Cj1293/Cj1294, as well as the C. jejuni Pgl pair Cj1120c/Cj1121c using His6-tagged purified derivatives. The metabolites produced by these enzymes were identified using NMR spectroscopy at 500 and/or 600 MHz with a cryogenically cooled probe for optimal sensitivity. The metabolites of Cj1293 (PseB) and HP0840 (FlaA1) were found to be labile and could only be characterized by NMR analysis directly in aqueous reaction buffer. The Cj1293 and HP0840 enzymes exhibited C6 dehydratase as well as a newly identified C5 epimerase activity that resulted in the production of both UDP-2-acetamido-2,6-dideoxy-
-L-arabino-4-hexulose and UDP-2-acetamido-2,6-dideoxy--D-xylo-4-hexulose. In contrast, the Pgl dehydratase Cj1120c (PglF) was found to possess only C6 dehydratase activity generating UDP-2-acetamido-2,6-dideoxy--D-xylo-4-hexulose. Substrate-specificity studies demonstrated that the flagellar aminotransferases HP0366 and Cj1294 utilize only UDP-2-acetamido-2,6-dideoxy--L-arabino-4-hexulose as substrate producing UDP-4-amino-4,6-dideoxy--L-AltNAc, a precursor in the Pse biosynthetic pathway. In contrast, the Pgl aminotransferase Cj1121c (PglE) utilizes only UDP-2-acetamido-2,6-dideoxy--D-xylo-4-hexulose producing UDP-4-amino-4,6-dideoxy--D-GlcNAc (UDP-2-acetamido-4-amino-2,4,6-trideoxy--D-glucopyranose), a precursor used in the production of the Pgl glycan component 2,4-diacetamido-2,4,6-trideoxy--D-glucopyranose.
Received for publication, October 11, 2005 , and in revised form, November 10, 2005.
* This study was supported by the National Research Council Genomics and Health Initiative. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1–S5 and supplemental Tables S1 and S2.
1 To whom correspondence should be addressed. Tel.: 613-990-0839; Fax: 613-952-9092; E-mail: susan.logan{at}nrc-cnrc.gc.ca.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  I. C Schoenhofen, E. Vinogradov, D. M Whitfield, J.-R. Brisson, and S. M Logan The CMP-legionaminic acid pathway in Campylobacter: Biosynthesis involving novel GDP-linked precursors Glycobiology, July 1, 2009; 19(7): 715 - 725. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. B. Tabei, P. G. Hitchen, M. J. Day-Williams, S. Merino, R. Vart, P.-C. Pang, G. J. Horsburgh, S. Viches, M. Wilhelms, J. M. Tomas, et al. An Aeromonas caviae Genomic Island Is Required for both O-Antigen Lipopolysaccharide Biosynthesis and Flagellin Glycosylation J. Bacteriol., April 15, 2009; 191(8): 2851 - 2863. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Chaban, S. M. Logan, J. F. Kelly, and K. F. Jarrell AglC and AglK Are Involved in Biosynthesis and Attachment of Diacetylated Glucuronic Acid to the N-Glycan in Methanococcus voltae J. Bacteriol., January 1, 2009; 191(1): 187 - 195. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. B. Olivier and B. Imperiali Crystal Structure and Catalytic Mechanism of PglD from Campylobacter jejuni J. Biol. Chem., October 10, 2008; 283(41): 27937 - 27946. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. D. Cook and H. M. Holden GDP-4-Keto-6-deoxy-D-mannose 3-Dehydratase, Accommodating a Sugar Substrate in the Active Site J. Biol. Chem., February 15, 2008; 283(7): 4295 - 4303. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Sharon Celebrating the golden anniversary of the discovery of bacillosamine, the diamino sugar of a Bacillus, Glycobiology, November 1, 2007; 17(11): 1150 - 1155. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Guerry, C. P. Ewing, I. C. Schoenhofen, and S. M. Logan Protein Glycosylation in Campylobacter jejuni: Partial Suppression of pglF by Mutation of pseC J. Bacteriol., September 15, 2007; 189(18): 6731 - 6733. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Chamot-Rooke, B. Rousseau, F. Lanternier, G. Mikaty, E. Mairey, C. Malosse, G. Bouchoux, V. Pelicic, L. Camoin, X. Nassif, et al. Alternative Neisseria spp. type IV pilin glycosylation with a glyceramido acetamido trideoxyhexose residue PNAS, September 11, 2007; 104(37): 14783 - 14788. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. McNally, A. J. Aubry, J. P. M. Hui, N. H. Khieu, D. Whitfield, C. P. Ewing, P. Guerry, J.-R. Brisson, S. M. Logan, and E. C. Soo Targeted Metabolomics Analysis of Campylobacter coli VC167 Reveals Legionaminic Acid Derivatives as Novel Flagellar Glycans J. Biol. Chem., May 11, 2007; 282(19): 14463 - 14475. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Vijayakumar, A. Merkx-Jacques, D. B. Ratnayake, I. Gryski, R. K. Obhi, S. Houle, C. M. Dozois, and C. Creuzenet Cj1121c, a Novel UDP-4-keto-6-deoxy-GlcNAc C-4 Aminotransferase Essential for Protein Glycosylation and Virulence in Campylobacter jejuni J. Biol. Chem., September 22, 2006; 281(38): 27733 - 27743. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. C. Schoenhofen, D. J. McNally, J.-R. Brisson, and S. M. Logan Elucidation of the CMP-pseudaminic acid pathway in Helicobacter pylori: synthesis from UDP-N-acetylglucosamine by a single enzymatic reaction Glycobiology, September 1, 2006; 16(9): 8C - 14C. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Liu and M. E. Tanner PseG of Pseudaminic Acid Biosynthesis: A UDP-SUGAR HYDROLASE AS A MASKED GLYCOSYLTRANSFERASE J. Biol. Chem., July 28, 2006; 281(30): 20902 - 20909. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. McNally, J. P. M. Hui, A. J. Aubry, K. K. K. Mui, P. Guerry, J.-R. Brisson, S. M. Logan, and E. C. Soo Functional Characterization of the Flagellar Glycosylation Locus in Campylobacter jejuni 81-176 Using a Focused Metabolomics Approach J. Biol. Chem., July 7, 2006; 281(27): 18489 - 18498. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. G. Hitchen and A. Dell Bacterial glycoproteomics Microbiology, June 1, 2006; 152(6): 1575 - 1580. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Weerapana and B. Imperiali Asparagine-linked protein glycosylation: from eukaryotic to prokaryotic systems Glycobiology, June 1, 2006; 16(6): 91R - 101R. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. Logan Flagellar glycosylation - a new component of the motility repertoire? Microbiology, May 1, 2006; 152(Pt 5): 1249 - 1262. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. C. Schoenhofen, V. V. Lunin, J.-P. Julien, Y. Li, E. Ajamian, A. Matte, M. Cygler, J.-R. Brisson, A. Aubry, S. M. Logan, et al. Structural and Functional Characterization of PseC, an Aminotransferase Involved in the Biosynthesis of Pseudaminic Acid, an Essential Flagellar Modification in Helicobacter pylori J. Biol. Chem., March 31, 2006; 281(13): 8907 - 8916. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |