Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis.
Sulfolipid-1 (SL-1) is an abundant sulfated glycolipid and potential virulence factor found in Mycobacterium tuberculosis. SL-1 consists of a trehalose-2-sulfate (T2S) disaccharide elaborated with four lipids. We identified and characterized a conserved mycobacterial sulfotransferase, Stf0, which generates the T2S moiety of SL-1. Biochemical studies demonstrated that the enzyme requires unmodified trehalose as substrate and is sensitive to small structural perturbations of the disaccharide. Disruption of stf0 in Mycobacterium smegmatis and M. tuberculosis resulted in the loss of T2S and SL-1 formation, respectively. The structure of Stf0 at a resolution of 2.6 A reveals the molecular basis of trehalose recognition and a unique dimer configuration that encloses the substrate into a bipartite active site. These data provide strong evidence that Stf0 carries out the first committed step in the biosynthesis of SL-1 and establish a system for probing the role of SL-1 in M. tuberculosis infection.
Mougous
Joseph D
JD
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
Petzold
Christopher J
CJ
Senaratne
Ryan H
RH
Lee
Dong H
DH
Akey
David L
DL
Lin
Fiona L
FL
Munchel
Sarah E
SE
Pratt
Matthew R
MR
Riley
Lee W
LW
Leary
Julie A
JA
Berger
James M
JM
Bertozzi
Carolyn R
CR
D001665:Binding Sites; D002855:Chromatography, Thin Layer; D018360:Crystallography, X-Ray; D003600:Cytosol / Q000201:enzymology; D019992:Databases as Topic; D003720:Densitometry; D019281:Dimerization; D004187:Disaccharides / Q000737:chemistry; D006860:Hydrogen Bonding; D007700:Kinetics; D008055:Lipids / Q000096:biosynthesis* / Q000737:chemistry; D013058:Mass Spectrometry; D008956:Models, Chemical; D008958:Models, Molecular; D009161:Mycobacterium / Q000201:enzymology; D020102:Mycobacterium smegmatis / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D016366:Open Reading Frames; D010100:Oxygen / Q000737:chemistry; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012694:Serine / Q000737:chemistry; D013329:Structure-Activity Relationship; D015238:Sulfotransferases / Q000737:chemistry*; D019076:Transgenes; D014199:Trehalose / Q000737:chemistry; D014965:X-Rays
D004187:Disaccharides; D008055:Lipids; C015518:sulfolipids; D012694:Serine; D014199:Trehalose; D015238:Sulfotransferases; D010100:Oxygen