title: Characterization of the N-acetyl-α-D-glucosaminyl l-malate synthase and deacetylase functions for bacillithiol biosynthesis in Bacillus anthracis .
abstract: Bacillithiol (Cys-GlcN-malate, BSH) has recently been identified as a novel low-molecular weight thiol in Bacillus anthracis, Staphylococcus aureus, and several other Gram-positive bacteria lacking glutathione and mycothiol. We have now characterized the first two enzymes for the BSH biosynthetic pathway in B. anthracis, which combine to produce α-d-glucosaminyl l-malate (GlcN-malate) from UDP-GlcNAc and l-malate. The structure of the GlcNAc-malate intermediate has been determined, as have the kinetic parameters for the BaBshA glycosyltransferase (→GlcNAc-malate) and the BaBshB deacetylase (→GlcN-malate). BSH is one of only two natural products reported to contain a malyl glycoside, and the crystal structure of the BaBshA-UDP-malate ternary complex, determined in this work at 3.3 Å resolution, identifies several active-site interactions important for the specific recognition of l-malate, but not other α-hydroxy acids, as the acceptor substrate. In sharp contrast to the structures reported for the GlcNAc-1-d-myo-inositol-3-phosphate synthase (MshA) apo and ternary complex forms, there is no major conformational change observed in the structures of the corresponding BaBshA forms. A mutant strain of B. anthracis deficient in the BshA glycosyltransferase fails to produce BSH, as predicted. This B. anthracis bshA locus (BA1558) has been identified in a transposon-site hybridization study as required for growth, sporulation, or germination [Day, W. A., Jr., Rasmussen, S. L., Carpenter, B. M., Peterson, S. N., and Friedlander, A. M. (2007) J. Bacteriol. 189, 3296-3301], suggesting that the biosynthesis of BSH could represent a target for the development of novel antimicrobials with broad-spectrum activity against Gram-positive pathogens like B. anthracis. The metabolites that function in thiol redox buffering and homeostasis in Bacillus are not well understood, and we present a composite picture based on this and other recent work.
authors.affiliation: Center for Structural Biology, Wake Forest University School of Medicine,Winston-Salem, North Carolina 27157, USA.
authors.forename: Derek
authors.initials: D
authors.lastname: Parsonage
authors.forename: Gerald L
authors.initials: GL
authors.lastname: Newton
authors.forename: Robert C
authors.initials: RC
authors.lastname: Holder
authors.forename: Bret D
authors.initials: BD
authors.lastname: Wallace
authors.forename: Carleitta
authors.initials: C
authors.lastname: Paige
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Hamilton
authors.forename: Patricia C
authors.initials: PC
authors.lastname: Dos Santos
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Redinbo
authors.forename: Sean D
authors.initials: SD
authors.lastname: Reid
authors.forename: Al
authors.initials: A
authors.lastname: Claiborne
mesh_terms: D001408:Bacillus anthracis / Q000201:enzymology* / Q000378:metabolism; D001665:Binding Sites; D001894:Borohydrides; D003545:Cysteine / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism*; D005944:Glucosamine / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000378:metabolism; D006020:Glycopeptides; D016695:Glycosyltransferases / Q000096:biosynthesis / Q000378:metabolism; D007294:Inositol; D019753:Intramolecular Lyases; D008970:Molecular Weight; D010084:Oxidation-Reduction; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014530:Uridine Diphosphate / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D001894:Borohydrides; D006020:Glycopeptides; D013438:Sulfhydryl Compounds; C543521:bacillithiol; C089265:mycothiol; C014651:mercaptoundecahydrododecaborate; D007294:Inositol; D014530:Uridine Diphosphate; D016695:Glycosyltransferases; D019753:Intramolecular Lyases; C105497:D-myo-inositol-3-phosphate synthase; D003545:Cysteine; D005944:Glucosamine
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