Covalent CouN7 Enzyme Intermediate for Acyl Group Shuttling in Aminocoumarin Biosynthesis

Carl J. Balibar; Sylvie Garneau-Tsodikova; Christopher T. Walsh

doi:10.1016/j.chembiol.2007.05.007

Covalent CouN7 Enzyme Intermediate for Acyl Group Shuttling in Aminocoumarin Biosynthesis

Carl J. Balibar, Sylvie Garneau-Tsodikova, Christopher T. Walsh

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

The last stages of assembly of the aminocoumarin antibiotics, clorobiocin and coumermycin A₁, which target the GyrB subunits of bacterial DNA gyrase, involve enzymatic transfer of the pyrrolyl-2-carbonyl acyl group from a carrier protein (CloN1/CouN1) to the 3′-OH of the noviosyl moiety of the antibiotic scaffold. The enzyme, CouN7, will catalyze both the forward and back reaction on both arms of the coumermycin scaffold. This occurs via an O-acyl-Ser₁₀₁-CouN7 intermediate, as shown by transient labeling of the enzyme with [¹⁴C]acetyl-S-CouN1 as donor and by inactivating mutation of the active site, Ser₁₀₁, to Ala. The intermediacy of the pyrrolyl-2-carbonyl-O-CouN7 allows net pyrrole transfer between distinct aminocoumarin scaffolds, for example, between the descarbamoylnovobiocin scaffold and coumermycin A₁ and vice versa. CouN7 also allows shuttling of surrogate acyl groups between noviosyl-aminocoumarin scaffolds to generate new antibiotic variants.

Original language	English
Pages (from-to)	679-690
Number of pages	12
Journal	Chemistry and Biology
Volume	14
Issue number	6
DOIs	https://doi.org/10.1016/j.chembiol.2007.05.007
State	Published - Jun 25 2007

Bibliographical note

Funding Information:
We thank Dr. Micha Fridman for the gift of 5-methylthiophene-CoA and Dr. Danica Galonic for a careful reading of the manuscript. Supported in part by National Institutes of Health grant GM 20011 (C.T.W.) and a Department of Defense National Defense Science and Engineering Graduate Fellowship (C.J.B.).

Keywords

CHEMBIOL
MICROBIO

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

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10.1016/j.chembiol.2007.05.007

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title = "Covalent CouN7 Enzyme Intermediate for Acyl Group Shuttling in Aminocoumarin Biosynthesis",

abstract = "The last stages of assembly of the aminocoumarin antibiotics, clorobiocin and coumermycin A1, which target the GyrB subunits of bacterial DNA gyrase, involve enzymatic transfer of the pyrrolyl-2-carbonyl acyl group from a carrier protein (CloN1/CouN1) to the 3′-OH of the noviosyl moiety of the antibiotic scaffold. The enzyme, CouN7, will catalyze both the forward and back reaction on both arms of the coumermycin scaffold. This occurs via an O-acyl-Ser101-CouN7 intermediate, as shown by transient labeling of the enzyme with [14C]acetyl-S-CouN1 as donor and by inactivating mutation of the active site, Ser101, to Ala. The intermediacy of the pyrrolyl-2-carbonyl-O-CouN7 allows net pyrrole transfer between distinct aminocoumarin scaffolds, for example, between the descarbamoylnovobiocin scaffold and coumermycin A1 and vice versa. CouN7 also allows shuttling of surrogate acyl groups between noviosyl-aminocoumarin scaffolds to generate new antibiotic variants.",

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note = "Funding Information: We thank Dr. Micha Fridman for the gift of 5-methylthiophene-CoA and Dr. Danica Galonic for a careful reading of the manuscript. Supported in part by National Institutes of Health grant GM 20011 (C.T.W.) and a Department of Defense National Defense Science and Engineering Graduate Fellowship (C.J.B.).",

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AU - Walsh, Christopher T.

N1 - Funding Information: We thank Dr. Micha Fridman for the gift of 5-methylthiophene-CoA and Dr. Danica Galonic for a careful reading of the manuscript. Supported in part by National Institutes of Health grant GM 20011 (C.T.W.) and a Department of Defense National Defense Science and Engineering Graduate Fellowship (C.J.B.).

PY - 2007/6/25

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N2 - The last stages of assembly of the aminocoumarin antibiotics, clorobiocin and coumermycin A1, which target the GyrB subunits of bacterial DNA gyrase, involve enzymatic transfer of the pyrrolyl-2-carbonyl acyl group from a carrier protein (CloN1/CouN1) to the 3′-OH of the noviosyl moiety of the antibiotic scaffold. The enzyme, CouN7, will catalyze both the forward and back reaction on both arms of the coumermycin scaffold. This occurs via an O-acyl-Ser101-CouN7 intermediate, as shown by transient labeling of the enzyme with [14C]acetyl-S-CouN1 as donor and by inactivating mutation of the active site, Ser101, to Ala. The intermediacy of the pyrrolyl-2-carbonyl-O-CouN7 allows net pyrrole transfer between distinct aminocoumarin scaffolds, for example, between the descarbamoylnovobiocin scaffold and coumermycin A1 and vice versa. CouN7 also allows shuttling of surrogate acyl groups between noviosyl-aminocoumarin scaffolds to generate new antibiotic variants.

AB - The last stages of assembly of the aminocoumarin antibiotics, clorobiocin and coumermycin A1, which target the GyrB subunits of bacterial DNA gyrase, involve enzymatic transfer of the pyrrolyl-2-carbonyl acyl group from a carrier protein (CloN1/CouN1) to the 3′-OH of the noviosyl moiety of the antibiotic scaffold. The enzyme, CouN7, will catalyze both the forward and back reaction on both arms of the coumermycin scaffold. This occurs via an O-acyl-Ser101-CouN7 intermediate, as shown by transient labeling of the enzyme with [14C]acetyl-S-CouN1 as donor and by inactivating mutation of the active site, Ser101, to Ala. The intermediacy of the pyrrolyl-2-carbonyl-O-CouN7 allows net pyrrole transfer between distinct aminocoumarin scaffolds, for example, between the descarbamoylnovobiocin scaffold and coumermycin A1 and vice versa. CouN7 also allows shuttling of surrogate acyl groups between noviosyl-aminocoumarin scaffolds to generate new antibiotic variants.

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Covalent CouN7 Enzyme Intermediate for Acyl Group Shuttling in Aminocoumarin Biosynthesis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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