|Title||Mechanistic studies on PseB of pseudaminic acid biosynthesis: A UDP-N-acetylglucosamine 5-inverting 4,6-dehydratase|
|Publication Type||Journal Article|
|Year of Publication||2008|
|Authors||Morrison, JP, Schoenhofen, IC, Tanner, ME|
|Type of Article||Article|
|Keywords||6-DEHYDRATASE, Campylobacter jejuni, CAMPYLOBACTER-JEJUNI, dehydratase, DTDP-GLUCOSE 4, ENZYMES, FLAA1, FUNCTIONAL-CHARACTERIZATION, GDP-MANNOSE 4, HELICOBACTER-PYLORI, IDENTIFICATION, Inverting, MOTILITY, PATHWAY, PseB, Pseudaminic acid, UDP-N-acetylglucosamine 5-inverting 4|
UDP-N-acetylglucosamine 5-inverting 4,6-dehydratase (PseB) is a unique sugar nucleotide dehydratase that inverts the C-5 ’’ stereocentre during conversion of UDP-N-acetylglucosamine to UDP-2-acetamido2,6-dideoxy-beta-L-arabino-hexos-4-ulose. PseB catalyzes the first step in the biosynthesis of pseudaminic acid, which is found as a post-translational modification on the flagellin of Campylobacter jejuni and Helicobacter pylori. PseB is proposed to use its tightly bound NADP(+) to oxidize UDP-GlcNAc at C-4 ’’, enabling dehydration. The alpha,beta unsaturated ketone intermediate is then reduced by delivery of the hydride to C-6 ’’ and a proton to C-5 ’’. Consistent with this, PseB from C. jejuni has been found to incorporate deuterium into the C-5 ’’ position of product during catalysis in D2O. Likewise, PseB catalyzes solvent isotope exchange into the H-5 ’’ position of product, and eliminates HF from the alternate Substrate, UDP-6-deoxy-6-fluoro-GlcNAc. Mutants of the putative catalytic residues aspartate 126, lysine 127 and tyrosine 135 have severely compromised dehydratase, solvent isotope exchange, and HF elimination activities. (C) 2008 Elsevier Inc. All rights reserved.
|URL||<Go to ISI>://000261905800019|