Purification and characterization of a protein inhibitor of calcium-dependent proteases from rat liver

George N. Demartino, Dorothy E. Croall

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Soluble extracts of rat liver contain a protein inhibitor of calcium-dependent proteases. The inhibitor has an apparent Mr = 250,000 and is separated from the calcium-dependent proteases by gel-filtration chromatography in the presence of EGTA. The inhibitor has been purified by affinity chromatography using a calcium-dependent protease covalently linked to Affi-Gel 15. The inhibitor specifically binds to this affinity resin in a calcium-dependent manner and elutes in the presence of EDTA or EGTA. The purified inhibitor appears as a single protein with Mr = 125,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Presumably it is a dimer under nondenaturing conditions. The inhibitor inhibits each of two calcium-dependent proteases from rat liver and from other tissues and species. However, it has no effect on any other protease tested.

Original languageEnglish (US)
Pages (from-to)713-720
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume232
Issue number2
DOIs
StatePublished - Aug 1 1984

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Liver
Purification
Rats
Peptide Hydrolases
Calcium
Egtazic Acid
Proteins
Affinity chromatography
Liver Extracts
Chromatography
Electrophoresis
Affinity Chromatography
Edetic Acid
Sodium Dodecyl Sulfate
Dimers
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Resins
Gels
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification and characterization of a protein inhibitor of calcium-dependent proteases from rat liver. / Demartino, George N.; Croall, Dorothy E.

In: Archives of Biochemistry and Biophysics, Vol. 232, No. 2, 01.08.1984, p. 713-720.

Research output: Contribution to journalArticle

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