Sensitive fluorometric determination of plasminogen activator in cell lysates and supernatants

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Abstract
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.