X‐ray crystal structure of rabbit N‐acetylglucosaminyltransferase I: catalytic mechanism and a new protein superfamily

N‐acetylglucosaminyltransferase I (GnT I) serves as the gateway from oligomannose to hybrid and complex N‐glycans and plays a critical role in mammalian development and possibly all metazoans. We have determined the X‐ray crystal structure of the catalytic fragment of GnT I in the absence and presence of bound UDP‐GlcNAc/Mn2+ at 1.5 and 1.8 Å resolution, respectively. The structures identify residues critical for substrate binding and catalysis and provide evidence for similarity, at the mechanistic level, to the deglycosylation step of retaining β‐glycosidases. The structuring of a 13 residue loop, resulting from UDP‐GlcNAc/Mn2+ binding, provides an explanation for the ordered sequential ‘Bi Bi’ kinetics shown by GnT I. Analysis reveals a domain shared with Bacillus subtilis glycosyltransferase SpsA, bovine β‐1,4‐galactosyl transferase 1 and Escherichia coli N‐acetylglucosamine‐1‐phosphate uridyltransferase. The low sequence identity, conserved fold and related functional features shown by this domain define a superfamily whose members probably share a common ancestor. Sequence analysis and protein threading show that the domain is represented in proteins from several glycosyltransferase families.

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