Sensitive, coupled assay for plasminogen activator using a thiol ester substrate for plasmin

Conference · Tue Jan 01 00:00:00 EST 1980

OSTI ID:6946467

Coleman, P L; Green, G D.J.

Several assays for plasminogen activator employ a direct assay method. These are remarkably sensitive methods, yet they suffer in comparison to the sensitivity of coupled methods. Coupling the assay with plasminogen not only amplifies the sensitivity by the multiplicative effect of plasmin, but insures that only those proteases specific for plasminogen are assayed. The choice of substrate for plasmin is critical. A thiol ester substrate, thiobenzyl benzyloxy-carbonyl-L-lysinate (Z-Lys-SBzl), has been synthesized which combines high k/sub cat/ with alkaline stability. In an effort to characterize the plasminogen activator from hepatoma tissue culture (HTC) and its hormonally-controlled inhibitor, Z-Lys-SBzl was used in a coupled approach providing an assay which is superior to the /sup 125/I-fibrinolytic assay. It is also extremely sensitive to plasminogen activator and can be used for routine analysis of purification as well as kinetic and binding studies. (ERB)

• Research Organization: Michigan Univ., Ann Arbor (USA). Dept. of Human Genetics; Brookhaven National Lab., Upton, NY (USA)
• DOE Contract Number: AC02-76CH00016
• OSTI ID: 6946467
• Report Number(s): BNL-28298; CONF-8005114-1
• Resource Relation: Conference: Contributions to hemostasis meeting, Detroit, MI, USA, 19 May 1980
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
BIOASSAY
SENSITIVITY
SPECIFICITY
BLOOD COAGULATION FACTORS
ENZYME ACTIVITY
BIOCHEMICAL REACTION KINETICS
CARBONIC ACID ESTERS
CELL CULTURES
ENZYME INHIBITORS
HEPATOMAS
SUBSTRATES
THIOLS
DISEASES
ESTERS
KINETICS
NEOPLASMS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
REACTION KINETICS
550200* - Biochemistry
550600 - Medicine
551000 - Physiological Systems