Crystal structure of a UDP-glucose-specific glycosyltransferase from a Mycobacterium species.
Glycosyltransferases (GTs) are a large and ubiquitous family of enzymes that specifically transfer sugar moieties to a range of substrates. Mycobacterium tuberculosis contains a large number of GTs, many of which are implicated in cell wall synthesis, yet the majority of these GTs remain poorly characterized. Here, we report the high resolution crystal structures of an essential GT (MAP2569c) from Mycobacterium avium subsp. paratuberculosis (a close homologue of Rv1208 from M. tuberculosis) in its apo- and ligand-bound forms. The structure adopted the GT-A fold and possessed the characteristic DXD motif that coordinated an Mn(2+) ion. Atypical of most GTs characterized to date, MAP2569c exhibited specificity toward the donor substrate, UDP-glucose. The structure of this ligated complex revealed an induced fit binding mechanism and provided a basis for this unique specificity. Collectively, the structural features suggested that MAP2569c may adopt a "retaining" enzymatic mechanism, which has implications for the classification of other GTs in this large superfamily.
Fulton
Zara
Z
Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800; Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800.
McAlister
Adrian
A
Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800.
Wilce
Matthew C J
MCJ
Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800.
Brammananth
Rajini
R
Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800; Department of Microbiology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800.
Zaker-Tabrizi
Leyla
L
Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800; Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800.
Perugini
Matthew A
MA
Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria 3010.
Bottomley
Stephen P
SP
Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800, Australia.
Coppel
Ross L
RL
Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800; Department of Microbiology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800.
Crellin
Paul K
PK
Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800; Department of Microbiology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800.
Rossjohn
Jamie
J
Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800; Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800. Electronic address: jamie.rossjohn@med.monash.edu.au.
Beddoe
Travis
T
Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800; Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, Clayton, Victoria 3800. Electronic address: travis.beddoe@med.monash.edu.au.
D020816:Amino Acid Motifs / Q000502:physiology; D001426:Bacterial Proteins / Q000737:chemistry*; D002473:Cell Wall / Q000201:enzymology*; D018360:Crystallography, X-Ray; D016695:Glycosyltransferases / Q000737:chemistry* / Q000145:classification; D008345:Manganese / Q000737:chemistry*; D009161:Mycobacterium / Q000201:enzymology*; D013379:Substrate Specificity; D014532:Uridine Diphosphate Glucose / Q000737:chemistry*
D001426:Bacterial Proteins; D008345:Manganese; D016695:Glycosyltransferases; D014532:Uridine Diphosphate Glucose