Crystal structure of mycothiol synthase (Rv0819) from Mycobacterium tuberculosis shows structural homology to the GNAT family of N-acetyltransferases

Matthew W. Vetting, Steven L. Roderick, Michael Yu, John S. Blanchard

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Mycothiol is the predominant low-molecular weight thiol produced by actinomycetes, including Mycobacterium tuberculosis. The last reaction in the biosynthetic pathway for mycothiol is catalyzed by mycothiol synthase (MshD), which acetylates the cysteinyl amine of cysteine-glucosamine-inositol (Cys-GlcN-Ins). The crystal structure of MshD was determined in the presence of coenzyme A and acetyl-CoA. MshD consists of two tandem-repeated domains, each exhibiting the Gcn5-related N-acetyltransferase (GNAT) fold. These two domains superimpose with a root-mean-square deviation of 1.7 Å over 88 residues, and each was found to bind one molecule of coenzyme, although the binding sites are quite different. The C-terminal domain has a similar active site to many GNAT members in which the acetyl group of the coenzyme is presented to an open active site slot. However, acetyl-CoA bound to the N-terminal domain is buried, and is apparently not positioned to promote acetyl transfer. A modeled substrate complex indicates that Cys-GlcN-Ins would only fill a portion of a negatively charged channel located between the two domains. This is the first structure determined for an enzyme involved in the biosynthesis of mycothiol.

Original languageEnglish (US)
Pages (from-to)1954-1959
Number of pages6
JournalProtein Science
Volume12
Issue number9
DOIs
StatePublished - Sep 1 2003

Fingerprint

Acetyltransferases
Mycobacterium tuberculosis
Acetyl Coenzyme A
Crystal structure
Glucosamine
Coenzymes
Inositol
Cysteine
Catalytic Domain
Actinobacteria
Biosynthesis
Biosynthetic Pathways
Coenzyme A
Sulfhydryl Compounds
Amines
Molecular Weight
Molecular weight
Binding Sites
Molecules
Substrates

Keywords

  • Acetyltransferase
  • GNAT fold
  • Mycothiol biosynthesis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystal structure of mycothiol synthase (Rv0819) from Mycobacterium tuberculosis shows structural homology to the GNAT family of N-acetyltransferases. / Vetting, Matthew W.; Roderick, Steven L.; Yu, Michael; Blanchard, John S.

In: Protein Science, Vol. 12, No. 9, 01.09.2003, p. 1954-1959.

Research output: Contribution to journalArticle

@article{0f7f7536719c4e6e93339d2dd8948c87,
title = "Crystal structure of mycothiol synthase (Rv0819) from Mycobacterium tuberculosis shows structural homology to the GNAT family of N-acetyltransferases",
abstract = "Mycothiol is the predominant low-molecular weight thiol produced by actinomycetes, including Mycobacterium tuberculosis. The last reaction in the biosynthetic pathway for mycothiol is catalyzed by mycothiol synthase (MshD), which acetylates the cysteinyl amine of cysteine-glucosamine-inositol (Cys-GlcN-Ins). The crystal structure of MshD was determined in the presence of coenzyme A and acetyl-CoA. MshD consists of two tandem-repeated domains, each exhibiting the Gcn5-related N-acetyltransferase (GNAT) fold. These two domains superimpose with a root-mean-square deviation of 1.7 {\AA} over 88 residues, and each was found to bind one molecule of coenzyme, although the binding sites are quite different. The C-terminal domain has a similar active site to many GNAT members in which the acetyl group of the coenzyme is presented to an open active site slot. However, acetyl-CoA bound to the N-terminal domain is buried, and is apparently not positioned to promote acetyl transfer. A modeled substrate complex indicates that Cys-GlcN-Ins would only fill a portion of a negatively charged channel located between the two domains. This is the first structure determined for an enzyme involved in the biosynthesis of mycothiol.",
keywords = "Acetyltransferase, GNAT fold, Mycothiol biosynthesis",
author = "Vetting, {Matthew W.} and Roderick, {Steven L.} and Michael Yu and Blanchard, {John S.}",
year = "2003",
month = "9",
day = "1",
doi = "10.1110/ps.03153703",
language = "English (US)",
volume = "12",
pages = "1954--1959",
journal = "Protein Science",
issn = "0961-8368",
publisher = "Cold Spring Harbor Laboratory Press",
number = "9",

}

TY - JOUR

T1 - Crystal structure of mycothiol synthase (Rv0819) from Mycobacterium tuberculosis shows structural homology to the GNAT family of N-acetyltransferases

AU - Vetting, Matthew W.

AU - Roderick, Steven L.

AU - Yu, Michael

AU - Blanchard, John S.

PY - 2003/9/1

Y1 - 2003/9/1

N2 - Mycothiol is the predominant low-molecular weight thiol produced by actinomycetes, including Mycobacterium tuberculosis. The last reaction in the biosynthetic pathway for mycothiol is catalyzed by mycothiol synthase (MshD), which acetylates the cysteinyl amine of cysteine-glucosamine-inositol (Cys-GlcN-Ins). The crystal structure of MshD was determined in the presence of coenzyme A and acetyl-CoA. MshD consists of two tandem-repeated domains, each exhibiting the Gcn5-related N-acetyltransferase (GNAT) fold. These two domains superimpose with a root-mean-square deviation of 1.7 Å over 88 residues, and each was found to bind one molecule of coenzyme, although the binding sites are quite different. The C-terminal domain has a similar active site to many GNAT members in which the acetyl group of the coenzyme is presented to an open active site slot. However, acetyl-CoA bound to the N-terminal domain is buried, and is apparently not positioned to promote acetyl transfer. A modeled substrate complex indicates that Cys-GlcN-Ins would only fill a portion of a negatively charged channel located between the two domains. This is the first structure determined for an enzyme involved in the biosynthesis of mycothiol.

AB - Mycothiol is the predominant low-molecular weight thiol produced by actinomycetes, including Mycobacterium tuberculosis. The last reaction in the biosynthetic pathway for mycothiol is catalyzed by mycothiol synthase (MshD), which acetylates the cysteinyl amine of cysteine-glucosamine-inositol (Cys-GlcN-Ins). The crystal structure of MshD was determined in the presence of coenzyme A and acetyl-CoA. MshD consists of two tandem-repeated domains, each exhibiting the Gcn5-related N-acetyltransferase (GNAT) fold. These two domains superimpose with a root-mean-square deviation of 1.7 Å over 88 residues, and each was found to bind one molecule of coenzyme, although the binding sites are quite different. The C-terminal domain has a similar active site to many GNAT members in which the acetyl group of the coenzyme is presented to an open active site slot. However, acetyl-CoA bound to the N-terminal domain is buried, and is apparently not positioned to promote acetyl transfer. A modeled substrate complex indicates that Cys-GlcN-Ins would only fill a portion of a negatively charged channel located between the two domains. This is the first structure determined for an enzyme involved in the biosynthesis of mycothiol.

KW - Acetyltransferase

KW - GNAT fold

KW - Mycothiol biosynthesis

UR - http://www.scopus.com/inward/record.url?scp=0042511920&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0042511920&partnerID=8YFLogxK

U2 - 10.1110/ps.03153703

DO - 10.1110/ps.03153703

M3 - Article

C2 - 12930994

AN - SCOPUS:0042511920

VL - 12

SP - 1954

EP - 1959

JO - Protein Science

JF - Protein Science

SN - 0961-8368

IS - 9

ER -