Calcium-dependent affinity purification of transglutaminase from rat liver cytosol.
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 5 mM calcium the enzyme binds to casein Sepharose and is subsequently eluted with 5 mM 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.
Croall
D E
DE
DeMartino
G N
GN
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002154:Calpain / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000302:isolation & purification; D051381:Rats; D011503:Transglutaminases / Q000302:isolation & purification* / Q000378:metabolism
D011503:Transglutaminases; D010447:Peptide Hydrolases; D002154:Calpain; D002118:Calcium