| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J Biol Chem, Vol. 274, Issue 44, 31391-31400, October 29, 1999
,
From the The lipopolysaccharide of Haemophilus
influenzae contains a single
3-deoxy-D-manno-octulosonic acid (Kdo) residue
derivatized with either a phosphate or an ethanolamine pyrophosphate
moiety at the 4-OH position. In previous studies, we identified a
kinase unique to H. influenzae extracts that phosphorylates
Kdo-lipid IVA, a key precursor of lipopolysaccharide in
this organism. We have now identified the gene encoding the Kdo kinase
by using an expression cloning approach. A cosmid library containing
random DNA fragments from H. influenzae strain Rd was
constructed in Escherichia coli. Extracts of 472 colonies
containing individual hybrid cosmids were assayed for Kdo kinase
activity. A single hybrid cosmid directing expression of the kinase was
found. The kinase gene was identified by activity assays, sub-cloning,
and DNA sequencing. When the putative kinase gene was expressed in E. coli behind a T7 promoter, massive overproduction of
kinase activity was achieved (~8000-fold higher than in H. influenzae membranes). The catalytic properties and the product
generated by the overexpressed kinase, assayed with Kdo-lipid
IVA as the substrate, were the same as observed with
H. influenzae membranes. Unexpectedly, the kinase gene was
identical to a previously characterized open reading frame
(orfZ), which had been shown to be important for
establishing bacteremia in an infant rat model (Hood, D. W., Deadman, M. E., Allen, T., Masoud, H., Martin, A.,
Brisson, J. R., Fleischmann, R., Venter, J. C., Richards,
J. C., and Moxon, E. R. (1996) Mol. Microbiol.
22, 951-965). However, based solely on the genome sequence of H. influenzae Rd, no biochemical function had been assigned to the
product of orfZ, which we now designate kdkA
("Kdo kinase A"). Although Kdo phosphorylation may be critical for
bacterial virulence of H. influenzae, it does not appear to be required for growth.
Department of Biochemistry, Duke University Medical
Center, Durham, North Carolina 27710 and § Middle Atlantic Mass
Spectrometry Laboratory, Department of Pharmacology and Molecular
Sciences, The Johns Hopkins University School of Medicine,
Baltimore, Maryland 21205-2185
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Harper, J. D. Boyce, A. D. Cox, F. St. Michael, I. W. Wilkie, P. J. Blackall, and B. Adler Pasteurella multocida Expresses Two Lipopolysaccharide Glycoforms Simultaneously, but Only a Single Form Is Required for Virulence: Identification of Two Acceptor-Specific Heptosyl I Transferases Infect. Immun., August 1, 2007; 75(8): 3885 - 3893. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Trent, C. M. Stead, A. X. Tran, and J. V. Hankins Invited review: Diversity of endotoxin and its impact on pathogenesis Innate Immunity, August 1, 2006; 12(4): 205 - 223. [Abstract] [PDF]
|
|
|
|
 |
|
 S. Gronow, W. Brabetz, B. Lindner, and H. Brade OpsX from Haemophilus influenzae Represents a Novel Type of Heptosyltransferase I in Lipopolysaccharide Biosynthesis J. Bacteriol., September 1, 2005; 187(17): 6242 - 6247. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. W. Hood, G. Randle, A. D. Cox, K. Makepeace, J. Li, E. K. H. Schweda, J. C. Richards, and E. R. Moxon Biosynthesis of Cryptic Lipopolysaccharide Glycoforms in Haemophilus influenzae Involves a Mechanism Similar to That Required for O-Antigen Synthesis J. Bacteriol., November 1, 2004; 186(21): 7429 - 7439. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. L. Horstman, S. J. Bauman, and M. J. Kuehn Lipopolysaccharide 3-Deoxy-D-manno-octulosonic Acid (Kdo) Core Determines Bacterial Association of Secreted Toxins J. Biol. Chem., February 27, 2004; 279(9): 8070 - 8075. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. E. Swords, P. A. Jones, and M. A. Apicella Review: The lipo-oligosaccharides of Haemophilus influenzae: an interesting array of characters Innate Immunity, June 1, 2003; 9(3): 131 - 144. [Abstract] [PDF]
|
|
|
|
|
|
L. Brade, S. Gronow, N. Wimmer, P. Kosma, and H. Brade Monoclonal antibodies against 3-deoxy-{alpha}-D-manno-oct-2-ulosonic acid (Kdo) and D-glycero-{alpha}-D-talo-oct-2-ulosonic acid (Ko) Innate Immunity, October 1, 2002; 8(5): 357 - 364. [Abstract] [PDF]
|
|
|
|
 |
|
 A. Krupa and N. Srinivasan Lipopolysaccharide phosphorylating enzymes encoded in the genomes of Gram-negative bacteria are related to the eukaryotic protein kinases Protein Sci., June 1, 2002; 11(6): 1580 - 1584. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. W. Hood, A. D. Cox, W. W. Wakarchuk, M. Schur, E. K.H. Schweda, S. L. Walsh, M. E. Deadman, A. Martin, E. R. Moxon, and J. C. Richards Genetic basis for expression of the major globotetraose-containing lipopolysaccharide from H. influenzae strain Rd (RM118) Glycobiology, November 1, 2001; 11(11): 957 - 967. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Gronow and H. Brade Invited review: Lipopolysaccharide biosynthesis: which steps do bacteria need to survive? Innate Immunity, February 1, 2001; 7(1): 3 - 23. [Abstract] [PDF]
|
|
|
|
|
|
W. Brabetz, S. Muller-Loennies, and H. Brade 3-Deoxy-D-manno-oct-2-ulosonic Acid (Kdo) Transferase (WaaA) and Kdo Kinase (KdkA) of Haemophilus influenzae Are Both Required to Complement a waaA Knockout Mutation of Escherichia coli J. Biol. Chem., November 3, 2000; 275(45): 34954 - 34962. [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 |
