Purification and properties of thioltransferase from monkey small intestinal mucosa

Its role in protein-s-thiolation

Outhiriaradjou Benard, K. A. Balasubramanian

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Modification of protein thiol by mixed disulfide formation with low molecular weight cellular thiols has been proposed as one of the post-translational modifications of amino acid side chains and is known to be catalyzed by thioltransferase. Intestinal mucosa is susceptible to oxidative injury and is likely to form protein mixed disulfide during oxidative stress. In the present study; thioltransferase was purified from monkey small intestinal mucosa and its role in protein-s-thiolation was investigated. The purified enzyme was homogeneous, as judged by polyacrylamide gel electrophoresis under reducing conditions. The enzyme, with a molecular weight of 52 kDa, was a monomeric protein, which showed optimum activity at pH 8.0 with hydroxyethyl disulfide as substrate. The enzyme specifically cleaved the disulfide bond of the synthetic substrate, hydroxyethyl disulfide, in the presence of reduced glutathione (GSH) with the formation of oxidized glutathione (GSSG) as shown by high performance liquid chromatography. The enzyme also catalyzed protein thiolation of monkey intestinal mitochondria when incubated with glutathione disulfide. These studies have shown that thioltransferase purified from intestinal mucosa could catalyze dethiolation and thiolation.

Original languageEnglish (US)
Pages (from-to)1051-1059
Number of pages9
JournalInternational Journal of Biochemistry and Cell Biology
Volume28
Issue number9
DOIs
StatePublished - Sep 1996
Externally publishedYes

Fingerprint

Glutaredoxins
Intestinal Mucosa
Disulfides
Haplorhini
Purification
Glutathione Disulfide
Enzymes
Proteins
Sulfhydryl Compounds
Molecular Weight
Molecular weight
Mitochondria
Oxidative stress
High performance liquid chromatography
Substrates
Post Translational Protein Processing
Electrophoresis
Glutathione
Polyacrylamide Gel Electrophoresis
Oxidative Stress

Keywords

  • Monkey intestinal mucosa
  • Protein-s-thiolation
  • Thiol-disulfide exchange activity
  • Thioltransferase

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

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title = "Purification and properties of thioltransferase from monkey small intestinal mucosa: Its role in protein-s-thiolation",
abstract = "Modification of protein thiol by mixed disulfide formation with low molecular weight cellular thiols has been proposed as one of the post-translational modifications of amino acid side chains and is known to be catalyzed by thioltransferase. Intestinal mucosa is susceptible to oxidative injury and is likely to form protein mixed disulfide during oxidative stress. In the present study; thioltransferase was purified from monkey small intestinal mucosa and its role in protein-s-thiolation was investigated. The purified enzyme was homogeneous, as judged by polyacrylamide gel electrophoresis under reducing conditions. The enzyme, with a molecular weight of 52 kDa, was a monomeric protein, which showed optimum activity at pH 8.0 with hydroxyethyl disulfide as substrate. The enzyme specifically cleaved the disulfide bond of the synthetic substrate, hydroxyethyl disulfide, in the presence of reduced glutathione (GSH) with the formation of oxidized glutathione (GSSG) as shown by high performance liquid chromatography. The enzyme also catalyzed protein thiolation of monkey intestinal mitochondria when incubated with glutathione disulfide. These studies have shown that thioltransferase purified from intestinal mucosa could catalyze dethiolation and thiolation.",
keywords = "Monkey intestinal mucosa, Protein-s-thiolation, Thiol-disulfide exchange activity, Thioltransferase",
author = "Outhiriaradjou Benard and Balasubramanian, {K. A.}",
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T2 - Its role in protein-s-thiolation

AU - Benard, Outhiriaradjou

AU - Balasubramanian, K. A.

PY - 1996/9

Y1 - 1996/9

N2 - Modification of protein thiol by mixed disulfide formation with low molecular weight cellular thiols has been proposed as one of the post-translational modifications of amino acid side chains and is known to be catalyzed by thioltransferase. Intestinal mucosa is susceptible to oxidative injury and is likely to form protein mixed disulfide during oxidative stress. In the present study; thioltransferase was purified from monkey small intestinal mucosa and its role in protein-s-thiolation was investigated. The purified enzyme was homogeneous, as judged by polyacrylamide gel electrophoresis under reducing conditions. The enzyme, with a molecular weight of 52 kDa, was a monomeric protein, which showed optimum activity at pH 8.0 with hydroxyethyl disulfide as substrate. The enzyme specifically cleaved the disulfide bond of the synthetic substrate, hydroxyethyl disulfide, in the presence of reduced glutathione (GSH) with the formation of oxidized glutathione (GSSG) as shown by high performance liquid chromatography. The enzyme also catalyzed protein thiolation of monkey intestinal mitochondria when incubated with glutathione disulfide. These studies have shown that thioltransferase purified from intestinal mucosa could catalyze dethiolation and thiolation.

AB - Modification of protein thiol by mixed disulfide formation with low molecular weight cellular thiols has been proposed as one of the post-translational modifications of amino acid side chains and is known to be catalyzed by thioltransferase. Intestinal mucosa is susceptible to oxidative injury and is likely to form protein mixed disulfide during oxidative stress. In the present study; thioltransferase was purified from monkey small intestinal mucosa and its role in protein-s-thiolation was investigated. The purified enzyme was homogeneous, as judged by polyacrylamide gel electrophoresis under reducing conditions. The enzyme, with a molecular weight of 52 kDa, was a monomeric protein, which showed optimum activity at pH 8.0 with hydroxyethyl disulfide as substrate. The enzyme specifically cleaved the disulfide bond of the synthetic substrate, hydroxyethyl disulfide, in the presence of reduced glutathione (GSH) with the formation of oxidized glutathione (GSSG) as shown by high performance liquid chromatography. The enzyme also catalyzed protein thiolation of monkey intestinal mitochondria when incubated with glutathione disulfide. These studies have shown that thioltransferase purified from intestinal mucosa could catalyze dethiolation and thiolation.

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