Use of synthetic oligosaccharide substrate analogs to map the active sites of N-acetylglucosaminyltransferases I and II.
Tables III and IV summarize substrate analog data presented in Tables I and II, respectively. Data for GlcNAc-T I shown in Tables I and III correlate very well with the crystal structure for GlcNAc-T I. This indicates that substitution of the various hydroxyl groups by hydrogen, [table: see text] O-methyl, and maybe even larger O-alkyl groups does not cause appreciable changes to either the overall conformation of the oligosaccharide or the binding mode, thus supporting this approach of chemical modification of oligosaccharide substrates for mapping of the binding site. There is as yet no crystal structure for GlcNAc-T II. These studies indicate both advantages and disadvantages of this approach for elucidating the catalytic and binding sites of an enzyme. Substrate analog data indicate which chemical groups in the substrate are essential for catalysis and binding and suggest the type of linkage involved (hydrogen bond donor or acceptor). However, no information has been obtained on the protein groups involved in these interactions. If a crystal structure is available, the substrate analog conclusions are primarily confirmatory. However, whether or not a crystal structure is available, this approach can be very helpful in the design of specific inhibitors.
Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G IX8, Canada.
Harry
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Schachter
Folkert
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Reck
Hans
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Paulsen
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D020134:Catalytic Domain; D002417:Cattle; D018360:Crystallography, X-Ray; D066298:In Vitro Techniques; D007700:Kinetics; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D009844:Oligosaccharides; D011134:Polysaccharides; D011994:Recombinant Proteins; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I; C063312:beta-1,3-galactosyl-0-glycosyl-glycoprotein beta-1,3-N-acetylglucosaminyltransferase