Cloning, expression, and biochemical characterization of a new histone deacetylase-like protein from Thermus caldophilus GK24

Young Mi Song, You Sun Kim, Dooil Kim, Dae Sil Lee, Ho Jeong Kwon

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Histone deactylases (HDACs) are members of an ancient enzyme family found in eukaryotes as well as in prokaryotes such as archaebacteria and eubacteria. We here report a new histone deacetylase (Tca HDAC) that was cloned from the genomic library of Thermus caldophilus GK24 based on homology analysis with human histone deacetylase1 (HDAC1). The gene contains an open reading frame encoding 375 amino acids with a calculated molecular mass of 42,188 Da and the deduced amino acid sequence of Tca HDAC showed a 31% homology to human HDAC1. The Tca HDAC gene was over-expressed in Escherichia coli using a Glutathione-S transferase (GST) fusion vector (pGEX-4T-1) and the purified protein showed a deacetylase activity toward the fluorogenic substrate for HDAC. Moreover, the enzyme activity was inhibited by trichostatin A, a specific HDAC inhibitor, in a dose-dependent manner. Optimum temperature and pH of the enzyme was found to be approximately 70 °C and 7.0, respectively. In addition, zinc ion is required for catalytic activity of the enzyme. Together, these data demonstrate that Tca HDAC is a new histone deacetylase-like enzyme from T. caldophilus GK24 and will be a useful tool for deciphering the role of HDAC in the prokaryote and development of new biochemical reactions.

Original languageEnglish
Pages (from-to)55-61
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume361
Issue number1
DOIs
Publication statusPublished - 2007 Sep 14

Fingerprint

Thermus
Histone Deacetylases
Cloning
Histones
Organism Cloning
Proteins
Enzymes
trichostatin A
Genes
Amino Acids
Genomic Library
Archaea
Enzyme activity
Molecular mass
Glutathione Transferase
Eukaryota
Fluorescent Dyes
Escherichia coli
Open Reading Frames
Zinc

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{f37113e5f714484db05c167c31166dba,
title = "Cloning, expression, and biochemical characterization of a new histone deacetylase-like protein from Thermus caldophilus GK24",
abstract = "Histone deactylases (HDACs) are members of an ancient enzyme family found in eukaryotes as well as in prokaryotes such as archaebacteria and eubacteria. We here report a new histone deacetylase (Tca HDAC) that was cloned from the genomic library of Thermus caldophilus GK24 based on homology analysis with human histone deacetylase1 (HDAC1). The gene contains an open reading frame encoding 375 amino acids with a calculated molecular mass of 42,188 Da and the deduced amino acid sequence of Tca HDAC showed a 31{\%} homology to human HDAC1. The Tca HDAC gene was over-expressed in Escherichia coli using a Glutathione-S transferase (GST) fusion vector (pGEX-4T-1) and the purified protein showed a deacetylase activity toward the fluorogenic substrate for HDAC. Moreover, the enzyme activity was inhibited by trichostatin A, a specific HDAC inhibitor, in a dose-dependent manner. Optimum temperature and pH of the enzyme was found to be approximately 70 °C and 7.0, respectively. In addition, zinc ion is required for catalytic activity of the enzyme. Together, these data demonstrate that Tca HDAC is a new histone deacetylase-like enzyme from T. caldophilus GK24 and will be a useful tool for deciphering the role of HDAC in the prokaryote and development of new biochemical reactions.",
author = "Song, {Young Mi} and Kim, {You Sun} and Dooil Kim and Lee, {Dae Sil} and Kwon, {Ho Jeong}",
year = "2007",
month = "9",
day = "14",
doi = "10.1016/j.bbrc.2007.06.147",
language = "English",
volume = "361",
pages = "55--61",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

Cloning, expression, and biochemical characterization of a new histone deacetylase-like protein from Thermus caldophilus GK24. / Song, Young Mi; Kim, You Sun; Kim, Dooil; Lee, Dae Sil; Kwon, Ho Jeong.

In: Biochemical and Biophysical Research Communications, Vol. 361, No. 1, 14.09.2007, p. 55-61.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cloning, expression, and biochemical characterization of a new histone deacetylase-like protein from Thermus caldophilus GK24

AU - Song, Young Mi

AU - Kim, You Sun

AU - Kim, Dooil

AU - Lee, Dae Sil

AU - Kwon, Ho Jeong

PY - 2007/9/14

Y1 - 2007/9/14

N2 - Histone deactylases (HDACs) are members of an ancient enzyme family found in eukaryotes as well as in prokaryotes such as archaebacteria and eubacteria. We here report a new histone deacetylase (Tca HDAC) that was cloned from the genomic library of Thermus caldophilus GK24 based on homology analysis with human histone deacetylase1 (HDAC1). The gene contains an open reading frame encoding 375 amino acids with a calculated molecular mass of 42,188 Da and the deduced amino acid sequence of Tca HDAC showed a 31% homology to human HDAC1. The Tca HDAC gene was over-expressed in Escherichia coli using a Glutathione-S transferase (GST) fusion vector (pGEX-4T-1) and the purified protein showed a deacetylase activity toward the fluorogenic substrate for HDAC. Moreover, the enzyme activity was inhibited by trichostatin A, a specific HDAC inhibitor, in a dose-dependent manner. Optimum temperature and pH of the enzyme was found to be approximately 70 °C and 7.0, respectively. In addition, zinc ion is required for catalytic activity of the enzyme. Together, these data demonstrate that Tca HDAC is a new histone deacetylase-like enzyme from T. caldophilus GK24 and will be a useful tool for deciphering the role of HDAC in the prokaryote and development of new biochemical reactions.

AB - Histone deactylases (HDACs) are members of an ancient enzyme family found in eukaryotes as well as in prokaryotes such as archaebacteria and eubacteria. We here report a new histone deacetylase (Tca HDAC) that was cloned from the genomic library of Thermus caldophilus GK24 based on homology analysis with human histone deacetylase1 (HDAC1). The gene contains an open reading frame encoding 375 amino acids with a calculated molecular mass of 42,188 Da and the deduced amino acid sequence of Tca HDAC showed a 31% homology to human HDAC1. The Tca HDAC gene was over-expressed in Escherichia coli using a Glutathione-S transferase (GST) fusion vector (pGEX-4T-1) and the purified protein showed a deacetylase activity toward the fluorogenic substrate for HDAC. Moreover, the enzyme activity was inhibited by trichostatin A, a specific HDAC inhibitor, in a dose-dependent manner. Optimum temperature and pH of the enzyme was found to be approximately 70 °C and 7.0, respectively. In addition, zinc ion is required for catalytic activity of the enzyme. Together, these data demonstrate that Tca HDAC is a new histone deacetylase-like enzyme from T. caldophilus GK24 and will be a useful tool for deciphering the role of HDAC in the prokaryote and development of new biochemical reactions.

UR - http://www.scopus.com/inward/record.url?scp=34547114742&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34547114742&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2007.06.147

DO - 10.1016/j.bbrc.2007.06.147

M3 - Article

VL - 361

SP - 55

EP - 61

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -