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Bioorg Med Chem. 1994 Nov;2(11):1231-41.

Key involvement of all three GlcNAc hydroxyl groups in the recognition of beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->6)-beta-D-Glcp-OR by N-acetylglucosaminyltransferase-V.

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1
Department of Chemistry, University of Alberta, Edmonton, Canada.

Abstract

N-Acetylglucosaminyltransferase-V (GlcNAc T-V) transfers a beta-linked GlcNAc residue from UDP-GlcNAc to OH-6' (of the alpha Man residue) in oligosaccharides terminating in the sequence beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->6)-beta-D-Glcp(or Manp)-OR (3, R = (CH2)7CH3). It was previously found that OH-4" (of the GlcNAc residue) in 3 was a critical element for substrate recognition by this enzyme. We show here that OH-3" and OH-6" are also key recognition elements. Four analogs of trisaccharide 3 where OH-3" and OH-6" were replaced, independently, by NH2 and NHAc groups, were prepared by multi-step chemical synthesis and kinetically evaluated as substrates for GlcNAc T-V from hamster kidney. These substitutions were selected since they replaced the OH groups with groups probing both hydrogen bonding and steric bulk. The 3"-modified compounds were found to be very poor substrates with Km values more than 50-fold elevated over that for 3 (26 microM) while the 6"-modified compounds were completely inactive. An intact 3,4,6 triol system in the terminal GlcNAc residue therefore appears to be the key polar group system that is recognized by this enzyme.

PMID:
7757419
[Indexed for MEDLINE]

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