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Sensitive fluorometric determination of plasminogen activator in cell lysates and supernatants
Author(s) -
Obrénovitch A.,
Maintier C.,
Maillet T.,
Mayer R.,
Kieda C.,
Monsigny M.
Publication year - 1983
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(83)80559-6
Subject(s) - plasmin , chemistry , plasminogen activator , fluorescence , tissue plasminogen activator , activator (genetics) , chromatography , microbiology and biotechnology , cell , substrate (aquarium) , biochemistry , enzyme , biology , receptor , ecology , physics , endocrinology , quantum mechanics
A fluorogenic substrate for plasmin, CBZ‐Gly—Pro—Arg‐AEC, has been synthesized and used to develop a new sensitive photometric and fluorometric assay of plasminogen activator activity. The fluorescence intensity of free AEC at 460 nm is about 3 orders of magnitude higher than that of acyl‐AEC. The release of AEC from the peptidyl derivative was monitored fluorometrically after extraction of free AEC in ethylacetate. Under such conditions, the K m was 0.16 mM. This method was used to monitor the activity of plasminogen activator synthetized by fibroblastic cells (BHK 21 C 13) either released in the supernantants or cell‐associated.