Crystal structure of vancosaminyltransferase GtfD from the vancomycin biosynthetic pathway: interactions with acceptor and nucleotide ligands.
The TDP-vancosaminyltransferase GtfD catalyzes the attachment of L-vancosamine to a monoglucosylated heptapeptide intermediate during the final stage of vancomycin biosynthesis. Glycosyltransferases from this and similar antibiotic pathways are potential tools for the design of new compounds that are effective against vancomycin resistant bacterial strains. We have determined the X-ray crystal structure of GtfD as a complex with TDP and the natural glycopeptide substrate at 2.0 A resolution. GtfD, a member of the bidomain GT-B glycosyltransferase superfamily, binds TDP in the interdomain cleft, while the aglycone acceptor binds in a deep crevice in the N-terminal domain. However, the two domains are more interdependent in terms of substrate binding and overall structure than was evident in the structures of closely related glycosyltransferases GtfA and GtfB. Structural and kinetic analyses support the identification of Asp13 as a catalytic general base, with a possible secondary role for Thr10. Several residues have also been identified as being involved in donor sugar binding and recognition.
Mulichak
Anne M
AM
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319, USA.
Lu
Wei
W
Losey
Heather C
HC
Walsh
Christopher T
CT
Garavito
R Michael
RM
D001665:Binding Sites; D002384:Catalysis; D003460:Crystallization; D018360:Crystallography, X-Ray; D005964:Glucosyltransferases / Q000737:chemistry*; D008024:Ligands; D009702:Nucleoside Diphosphate Sugars / Q000737:chemistry; D009711:Nucleotides / Q000737:chemistry*; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D013942:Thymine Nucleotides / Q000737:chemistry; D014640:Vancomycin / Q000096:biosynthesis* / Q000737:chemistry
D008024:Ligands; D009702:Nucleoside Diphosphate Sugars; D009711:Nucleotides; D013942:Thymine Nucleotides; C416082:thymine diphosphate epivancosamine; D014640:Vancomycin; D005964:Glucosyltransferases; C113848:glucosyltransferase D