Open AccessPurification and characterization of an inhibitor of plasminogen activator released by rat mammary adenocarcinoma cells
Author(s) -
GRANT Adrienne J.,
RAMSHAW Ian A.,
BADENOCHJONES Peter,
EICHNER Ronald D.,
HUNT Nicholas H.
Publication year - 1986
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1986.tb09445.x
Subject(s) - plasminogen activator , plasmin , urokinase , chromatofocusing , cell culture , microbiology and biotechnology , trypsin , isoelectric point , adenocarcinoma , chemistry , biology , biochemistry , endocrinology , enzyme , cancer , genetics
An inhibitor of plasminogen activator (PA) secreted by a tumorigenic, but non‐metastatic, rat mammary adenocarcinoma cell line has been purified to apparent homogeneity and characterized. It strongly inhibited human urokinase, but was 100 times less potent in inhibiting bovine trypsin and had no effect on plasmin or thrombin. A secreted, urokinase‐type PA ( M r 48000) and a cell‐associated PA from a metastatic rat adenocarcinoma cell line were also strongly inhibited. In contrast, a tissue‐type PA ( M r 66000), secreted by human melanoma cells, was only slightly inhibited. Purified inhibitor showed a band of M r 66000 in sodium dodecyl sulphate/polyacrylamide gel electrophoresis and an isoelectric point of 4.5 after chromatofocusing. The inhibition of human urokinase was non‐competitive.