Activity-based substrate profiling for Gcn5-related N-acetyltransferases: The use of chloroacetyl-coenzyme A to identify protein substrates

Michael Yu; Luiz Pedro Sorio De Carvalho; Guangxing Sun; John S. Blanchard

doi:10.1021/ja066298w

Activity-based substrate profiling for Gcn5-related N-acetyltransferases: The use of chloroacetyl-coenzyme A to identify protein substrates

Michael Yu, Luiz Pedro Sorio De Carvalho, Guangxing Sun, John S. Blanchard

Biochemistry

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

33 Scopus citations

Abstract

The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His₈) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the α-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes.

Original language	English (US)
Pages (from-to)	15356-15357
Number of pages	2
Journal	Journal of the American Chemical Society
Volume	128
Issue number	48
DOIs	https://doi.org/10.1021/ja066298w
State	Published - Dec 6 2006

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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title = "Activity-based substrate profiling for Gcn5-related N-acetyltransferases: The use of chloroacetyl-coenzyme A to identify protein substrates",

abstract = "The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His8) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the α-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes.",

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AU - De Carvalho, Luiz Pedro Sorio

AU - Sun, Guangxing

AU - Blanchard, John S.

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N2 - The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His8) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the α-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes.

AB - The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His8) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the α-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes.

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Activity-based substrate profiling for Gcn5-related N-acetyltransferases: The use of chloroacetyl-coenzyme A to identify protein substrates

Abstract

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