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

Producción científica: Article › revisión exhaustiva

17 Citas (Scopus)

Resumen

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.

Idioma original	English
Páginas (desde-hasta)	679-690
Número de páginas	12
Publicación	Chemistry and Biology
Volumen	14
N.º	6
DOI	https://doi.org/10.1016/j.chembiol.2007.05.007
Estado	Published - jun 25 2007

Nota bibliográfica

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.).

Financiación

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.).

Financiadores	Número del financiador
National Institutes of Health (NIH)
National Institute of General Medical Sciences DP2GM119177 Sophie Dumont National Institute of General Medical Sciences	R37GM020011
National Defense Science and Engineering Graduate

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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author = "Balibar, \{Carl J.\} and Sylvie Garneau-Tsodikova and Walsh, \{Christopher T.\}",

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

Resumen

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ASJC Scopus subject areas

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