Isolation and some molecular parameters of elastase-like normal proteinases from horse blood leucocytes.
Cytoplasmic granules were isolated from horse blood polymorphonuclear leucocytes by the heparin method and extracted with 0.9% NaCl by repeated freezing. Soluble proteins were separated on a column of Sephadex G-75 followed by chromatography on a column of CM-Sephadex with a NaCl gradient. Gel filtration, density-gradient centrifugation, isoelectric focusing and 0.1% sodium dodecyl sulphate/polyacrylamide-gel electrophoresis at pH 7.0 and at pH 4.5 were used to determine molecular parameters of proteinases. Three enzymes hydrolysing both casein and N-benzyloxycarbonyl-L-alanine nitrophenyl ester were found in the granule extract: proteinase 1, mol.wt. 38000, pI5.3; proteinase 2A, mol.wt. 24500, pI8.8; and proteinase 2B, mol.wt. 20500, pI above 10. The latter two elastase-like proteinases were purified to apparent homogeneity.
D000818:Animals; D002364:Caseins; D002553:Cerebroside-Sulfatase / Q000032:analysis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006736:Horses / Q000097:blood*; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007962:Leukocytes / Q000032:analysis / Q000201:enzymology*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000097:blood* / Q000302:isolation & purification; D011506:Proteins / Q000032:analysis; D014461:Ultracentrifugation
D002364:Caseins; D011506:Proteins; D002553:Cerebroside-Sulfatase; D010447:Peptide Hydrolases