Cibacron Blue-induced modification of neutral proteinase from horse blood leukocytes.
The proteolytic activity of the elastase-like proteinase from granules of horse blood leukocytes is retained on a column of Cibacron Blue-Sepharose and can be eluted with 0.5 M KSCN. During this procedure its mol. wt. is reduced from 49000 to 30000 and isoelectric point is shifted towards higher pH. The inactive protein not adsorbed on Cibacron Blue-Sepharose is strongly acidic and shows a mol. wt. of 20000. Upon mixing this protein with the modified enzyme the native proteinase is reconstituted as shown by polyacrylamide gel electrophoresis at pH 8.3 and isoelectric focusing in a sucrose gradient.
D000818:Animals; D000873:Anthracenes; D002846:Chromatography, Affinity; D003594:Cytoplasmic Granules / Q000201:enzymology; D006736:Horses / Q000097:blood*; D007526:Isoelectric Point; D007962:Leukocytes / Q000201:enzymology*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000097:blood*; D011134:Polysaccharides; D012685:Sepharose / Q000031:analogs & derivatives
D000873:Anthracenes; D011134:Polysaccharides; C017842:cibacron blue-sepharose; D012685:Sepharose; D010447:Peptide Hydrolases