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

Research output: Contribution to journalArticle

25 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)15356-15357
Number of pages2
JournalJournal of the American Chemical Society
Volume128
Issue number48
DOIs
StatePublished - Dec 6 2006

Fingerprint

Coenzymes
Acetyltransferases
Proteins
Histones
Substrates
Enzymes
Histone Acetyltransferases
Complex Mixtures
Sulfhydryl Compounds
chloroacetyl coenzyme A

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Activity-based substrate profiling for Gcn5-related N-acetyltransferases : The use of chloroacetyl-coenzyme A to identify protein substrates. / Yu, Michael; De Carvalho, Luiz Pedro Sorio; Sun, Guangxing; Blanchard, John S.

In: Journal of the American Chemical Society, Vol. 128, No. 48, 06.12.2006, p. 15356-15357.

Research output: Contribution to journalArticle

@article{dfded05438b847a893d314081475267f,
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.",
author = "Michael Yu and {De Carvalho}, {Luiz Pedro Sorio} and Guangxing Sun and Blanchard, {John S.}",
year = "2006",
month = "12",
day = "6",
doi = "10.1021/ja066298w",
language = "English (US)",
volume = "128",
pages = "15356--15357",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "48",

}

TY - JOUR

T1 - Activity-based substrate profiling for Gcn5-related N-acetyltransferases

T2 - The use of chloroacetyl-coenzyme A to identify protein substrates

AU - Yu, Michael

AU - De Carvalho, Luiz Pedro Sorio

AU - Sun, Guangxing

AU - Blanchard, John S.

PY - 2006/12/6

Y1 - 2006/12/6

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.

UR - http://www.scopus.com/inward/record.url?scp=33845292279&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845292279&partnerID=8YFLogxK

U2 - 10.1021/ja066298w

DO - 10.1021/ja066298w

M3 - Article

C2 - 17131985

AN - SCOPUS:33845292279

VL - 128

SP - 15356

EP - 15357

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 48

ER -