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Heterogeneity of human tissue‐type plasminogen activator
Author(s) -
Opdenakker Ghislain,
Bosman Fons,
Decock Benny,
Cabeza-Arvelaiz Yofre,
Van Damme Jo,
Billiau Alfons
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80241-2
Subject(s) - size exclusion chromatography , chemistry , polyclonal antibodies , plasminogen activator , microbiology and biotechnology , antibody , cell culture , biochemistry , chromatography , enzyme , biology , immunology , endocrinology , genetics
Tissue‐type plasminogen activator (t‐PA) from human melanoma cells (Bowes) was purified by immunosorbent chromatography on affinospecific polyclonal antibodies and gel filtration in the presence of KSCN. The immunosorbent eluate contained three major components of > 200, 85 and 65 kDa, respectively. The 65 kDa t‐PA component could be separated by gel filtration on Ultrogel AcA44 in the presence of KSCN to a pure preparation yielding a unique N‐terminal amino acid sequence. Immunoblot analysis, using affinospecific antibodies against t‐PA, was a specific and sensitive method to identify different types of t‐PA (I–IV), as well as t‐PA‐inhibitor complexes and degradation products in unstimulated melanoma cell culture fluids. Furthermore, the t‐PA preparations, produced by phorbol ester‐treated melanoma cells, were free of type IV and thus differed physicochemically from the constitutively produced t‐PA preparations. The composition of t‐PA from mammalian cell cultures is thus more complex than hitherto described.