Purification and characterization of a plasminogen activator secreted by a pig kidney cell line

M. Sudol, E. Reich

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11 Scopus citations

Abstract

The plasminogen activator secreted by calcitonin-treated pig kidney cells was purified, characterized and compared with human urinary urokinase. The purification procedure was based on the following steps: sulphopropyl-Sephadex chromatography, p-aminobenzamidine-Sepharose chromatography, preparative sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and isoelectrofocusing. The purified enzyme was obtained from the conditioned medium with a yield of 13% and a purification factor of 390-fold. Analysis by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis under non-reducing conditions showed one closely spaced doublet with an M(r) of 50000; in the presence of reducing agents, two additional bands of M(r) 30000 and 20000 appeared. The purified enzyme resembles the 53000-M(r) components of human urinary urokinase in amino acid composition and two-dimensional tryptic peptide maps and in its catalytic properties, and the two enzymes cross-react immunologically with rabbit antibodies raised against either. The enzyme appears to be different from tissue plasminogen activator secreted by HeLa cells.

Original languageEnglish
Pages (from-to)971-978
Number of pages8
JournalBiochemical Journal
Volume219
Issue number3
DOIs
StatePublished - 1984
Externally publishedYes

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