Crystal Structure of StaL, a Glycopeptide Antibiotic Sulfotransferase from Streptomyces toyocaensis

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1074/jbc.M61191200
AuthorSearch for: ; Search for: ; Search for: ; Search for: 1; Search for: ; Search for: 1; Search for: 1
Name affiliation
  1. National Research Council Canada. NRC Biotechnology Research Institute
FormatText
TypeArticle
Journal titleThe Journal of Biological Chemistry
ISSN0021-9258
1083-351X
Volume282
Issue17
Pages13073–13086
Subjectbiosynthesis; crystal structure; dimerization; fluorescence; in vitro; mutagenesis; pharmaceutical
Abstract
Over the past decade, antimicrobial resistance has emerged as a major public health crisis. Glycopeptide antibiotics such as vanco-mycin and teicoplanin are clinically important for the treatment of Gram-positive bacterial infections. StaL is a 3'-phosphoadenosine 5'-phosphosulfate-dependent sulfotransferase capable of sulfating the cross-linked heptapeptide substrate both in vivo and in vitro, yielding the product A47934, a unique teicoplanin-class glycopeptide antibiotic. The sulfonation reaction catalyzed by StaL constitutes the final step in A47934 biosynthesis. Here we report the crystal structure of StaL and its complex with the cofactor product 3'-phosphoadenosine 5'-phosphate. This is only the second prokaryotic sulfotransferase to be structurally characterized. StaL belongs to the large sulfotransferase family and shows higher similarity to cytosolic sulfotransferases (ST) than to the bacterial ST (Stf0). StaL has a novel dimerization motif, different from any other STs that have been structurally characterized. We have also applied molecular modeling to investigate the binding mode of the unique substrate, desulfo-A47934. Based on the structural analysis and modeling results, a series of residues was mutated and kinetically characterized. In addition to the conserved residues (Lys(12), His(67), and Ser(98)), molecular modeling, fluorescence quenching experiments, and mutagenesis studies identified several other residues essential for substrate binding and/or activity, including Trp(34), His(43), Phe(77), Trp(132), and Glu(205)
Publication date
PublisherAmerican Society for Biochemistry and Molecular Biology
LanguageEnglish
Peer reviewedYes
NRC numberNRC-BRI-49511
NPARC number3538904
Export citationExport as RIS
Report a correctionReport a correction
Record identifier251b36c1-0cfb-471d-8faa-d8e78500e190
Record created2009-03-01
Record modified2020-02-19
Report a problem on this page
Date modified: