Calcium-dependent affinity purification of transclutaminase from rat liver cytosol

Dorothy E. Croall, George N. DeMartino

Research output: Contribution to journalArticle

18 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)29-39
Number of pages11
JournalCell Calcium
Volume7
Issue number1
DOIs
StatePublished - 1986

Fingerprint

Cytosol
Caseins
Calcium
Sepharose
Transglutaminases
Liver
Affinity Chromatography
Liver Extracts
Egtazic Acid
Ion Exchange Chromatography
Enzymes
Transferases
Amines
Gel Chromatography
Chromatography
Peptide Hydrolases
Molecular Weight

ASJC Scopus subject areas

  • Cell Biology
  • Endocrinology

Cite this

Calcium-dependent affinity purification of transclutaminase from rat liver cytosol. / Croall, Dorothy E.; DeMartino, George N.

In: Cell Calcium, Vol. 7, No. 1, 1986, p. 29-39.

Research output: Contribution to journalArticle

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