Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis

Joseph D. Mougous; Christopher J. Petzold; Ryan H. Senaratne; Dong H. Lee; David L. Akey; Fiona L. Lin; Sarah E. Munchel; Matthew R. Pratt; Lee W. Riley; Julie A. Leary; James M. Berger; Carolyn R. Bertozzi

doi:10.1038/nsmb802

Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis

Joseph D. Mougous, Christopher J. Petzold, Ryan H. Senaratne, Dong H. Lee, David L. Akey, Fiona L. Lin, Sarah E. Munchel, Matthew R. Pratt, Lee W. Riley, Julie A. Leary, James M. Berger, Carolyn R. Bertozzi

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Abstract

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 Å 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.

Original language	English (US)
Pages (from-to)	721-729
Number of pages	9
Journal	Nature Structural and Molecular Biology
Volume	11
Issue number	8
DOIs	https://doi.org/10.1038/nsmb802
State	Published - Aug 2004
Externally published	Yes

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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Mougous, J. D., Petzold, C. J., Senaratne, R. H., Lee, D. H., Akey, D. L., Lin, F. L., Munchel, S. E., Pratt, M. R., Riley, L. W., Leary, J. A., Berger, J. M., & Bertozzi, C. R. (2004). Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis. Nature Structural and Molecular Biology, 11(8), 721-729. https://doi.org/10.1038/nsmb802

@article{3b9c9c0754454422a14f0c1e1cea22b4,

title = "Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis",

abstract = "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 {\AA} 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.",

author = "Mougous, {Joseph D.} and Petzold, {Christopher J.} and Senaratne, {Ryan H.} and Lee, {Dong H.} and Akey, {David L.} and Lin, {Fiona L.} and Munchel, {Sarah E.} and Pratt, {Matthew R.} and Riley, {Lee W.} and Leary, {Julie A.} and Berger, {James M.} and Bertozzi, {Carolyn R.}",

note = "Funding Information: We thank the TB Structural Genomics Consortium (http://www.doe-mbi.ucla. edu/TB) for partial funding. We are grateful to J. Holton and C. Ralston at the Advanced Light Source for their assistance with data collection and processing, D. Chen and M. Schelle for fruitful scientific discussions, members of the Berger and Bertozzi labs for careful review of the manuscript, D. Keating for sharing valuable sequence data and H. Schachman for use of equipment. J.D.M. was supported by a fellowship from the Ford Foundation. This work was supported by a grant from the US National Institutes of Health to C.R.B. (AI51622) and J.M.B. (P50-GM62410). Copyright: Copyright 2008 Elsevier B.V., All rights reserved.",

year = "2004",

month = aug,

doi = "10.1038/nsmb802",

language = "English (US)",

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T1 - Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis

AU - Mougous, Joseph D.

AU - Petzold, Christopher J.

AU - Senaratne, Ryan H.

AU - Lee, Dong H.

AU - Akey, David L.

AU - Lin, Fiona L.

AU - Munchel, Sarah E.

AU - Pratt, Matthew R.

AU - Riley, Lee W.

AU - Leary, Julie A.

AU - Berger, James M.

AU - Bertozzi, Carolyn R.

N1 - Funding Information: We thank the TB Structural Genomics Consortium (http://www.doe-mbi.ucla. edu/TB) for partial funding. We are grateful to J. Holton and C. Ralston at the Advanced Light Source for their assistance with data collection and processing, D. Chen and M. Schelle for fruitful scientific discussions, members of the Berger and Bertozzi labs for careful review of the manuscript, D. Keating for sharing valuable sequence data and H. Schachman for use of equipment. J.D.M. was supported by a fellowship from the Ford Foundation. This work was supported by a grant from the US National Institutes of Health to C.R.B. (AI51622) and J.M.B. (P50-GM62410). Copyright: Copyright 2008 Elsevier B.V., All rights reserved.

PY - 2004/8

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N2 - 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 Å 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.

AB - 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 Å 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.

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Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis

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