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 language | English |
|---|---|
| Pages (from-to) | 971-978 |
| Number of pages | 8 |
| Journal | Biochemical Journal |
| Volume | 219 |
| Issue number | 3 |
| DOIs | |
| State | Published - 1984 |
| Externally published | Yes |