Calcium-dependent affinity purification of transclutaminase from rat liver cytosol

Dorothy E. Croall; George N. DeMartino

doi:10.1016/0143-4160(86)90028-X

Calcium-dependent affinity purification of transclutaminase from rat liver cytosol

Dorothy E. Croall, George N. DeMartino

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Abstract

Tissue transglutaminase (E.C.2.3.2.13, R-glutaminyl-peptide: amine glutaminyl transferase), was purified from extracts of rat liver by calcium dependent affinity chromatography on casein-Sepharose. In the presence of 5mM calcium the enzyme binds to casein Sepharose and is subsequently eluted with 5mM EGTA. The enzyme has a molecular weight of 83,000 and its activity is dependent on calcium and reduced sulfhydryl residues. A widely distributed calcium-dependent protease (E.C. 3.4.22.17) copurified with transglutaminase by gel filtration and ion exchange chromatography. The separation of these activities prior to chromatography on casein-Sepharose is essential for the isolation of a stable transglutaminase by calcium-dependent affinity chromatography. Affinity chromatography using casein-Sepharose or other immobilized substrates may allow the calcium-dependent purification of a variety of transglutaminases.

Original language	English (US)
Pages (from-to)	29-39
Number of pages	11
Journal	Cell Calcium
Volume	7
Issue number	1
DOIs	https://doi.org/10.1016/0143-4160(86)90028-X
State	Published - Feb 1986

ASJC Scopus subject areas

Physiology
Molecular Biology
Cell Biology

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10.1016/0143-4160(86)90028-X

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title = "Calcium-dependent affinity purification of transclutaminase from rat liver cytosol",

abstract = "Tissue transglutaminase (E.C.2.3.2.13, R-glutaminyl-peptide: amine glutaminyl transferase), was purified from extracts of rat liver by calcium dependent affinity chromatography on casein-Sepharose. In the presence of 5mM calcium the enzyme binds to casein Sepharose and is subsequently eluted with 5mM EGTA. The enzyme has a molecular weight of 83,000 and its activity is dependent on calcium and reduced sulfhydryl residues. A widely distributed calcium-dependent protease (E.C. 3.4.22.17) copurified with transglutaminase by gel filtration and ion exchange chromatography. The separation of these activities prior to chromatography on casein-Sepharose is essential for the isolation of a stable transglutaminase by calcium-dependent affinity chromatography. Affinity chromatography using casein-Sepharose or other immobilized substrates may allow the calcium-dependent purification of a variety of transglutaminases.",

author = "Croall, {Dorothy E.} and DeMartino, {George N.}",

note = "Funding Information: grants from NIH of the American Heart and C. F. Graham Funding Information: This work was supported by G. N. D. and the Texas Affiliate D.E.C.. We thank J. Bower assistance.",

year = "1986",

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doi = "10.1016/0143-4160(86)90028-X",

language = "English (US)",

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T1 - Calcium-dependent affinity purification of transclutaminase from rat liver cytosol

AU - Croall, Dorothy E.

AU - DeMartino, George N.

N1 - Funding Information: grants from NIH of the American Heart and C. F. Graham Funding Information: This work was supported by G. N. D. and the Texas Affiliate D.E.C.. We thank J. Bower assistance.

PY - 1986/2

Y1 - 1986/2

N2 - Tissue transglutaminase (E.C.2.3.2.13, R-glutaminyl-peptide: amine glutaminyl transferase), was purified from extracts of rat liver by calcium dependent affinity chromatography on casein-Sepharose. In the presence of 5mM calcium the enzyme binds to casein Sepharose and is subsequently eluted with 5mM EGTA. The enzyme has a molecular weight of 83,000 and its activity is dependent on calcium and reduced sulfhydryl residues. A widely distributed calcium-dependent protease (E.C. 3.4.22.17) copurified with transglutaminase by gel filtration and ion exchange chromatography. The separation of these activities prior to chromatography on casein-Sepharose is essential for the isolation of a stable transglutaminase by calcium-dependent affinity chromatography. Affinity chromatography using casein-Sepharose or other immobilized substrates may allow the calcium-dependent purification of a variety of transglutaminases.

AB - Tissue transglutaminase (E.C.2.3.2.13, R-glutaminyl-peptide: amine glutaminyl transferase), was purified from extracts of rat liver by calcium dependent affinity chromatography on casein-Sepharose. In the presence of 5mM calcium the enzyme binds to casein Sepharose and is subsequently eluted with 5mM EGTA. The enzyme has a molecular weight of 83,000 and its activity is dependent on calcium and reduced sulfhydryl residues. A widely distributed calcium-dependent protease (E.C. 3.4.22.17) copurified with transglutaminase by gel filtration and ion exchange chromatography. The separation of these activities prior to chromatography on casein-Sepharose is essential for the isolation of a stable transglutaminase by calcium-dependent affinity chromatography. Affinity chromatography using casein-Sepharose or other immobilized substrates may allow the calcium-dependent purification of a variety of transglutaminases.

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Calcium-dependent affinity purification of transclutaminase from rat liver cytosol

Abstract

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