Human Plasminogen Variant Chicago III

Abstract

SummaryA new abnormal, variant, plasminogen Chicago III has been isolated from a patient with recurring deep vein thrombosis. Studies on the plasma fibrinolytic system of four family members showed no inheritance pattern. Kinetics of activation parameters of Chicago III plasminogen with different activators showed lowered catalytic rate constants from 159fold with urokinase, to 3fold with light (B) chain ‧ streptokinase complex, to 1.3fold with streptokinase. The Michaelis constants of activation of Chicago III plasminogen were 16fold higher with streptokinase, 6fold higher with light (B) chain ‧ streptokinase complex, and similar with urokinase. Each of the three activators exhibited different interaction characteristics with this variant zymogen, as they did with variant Chicago I and Chicago II plasminogens (previously reported). However, the latter variants were characterized by having normal catalytic rate constants of activation, with higher than normal apparent Michaelis constants of activation. Chicago III plasminogen, as well as Chicago I and II plasminogens, has a homogeneous population of molecules; the interpretation of the kinetic data was possible only in terms of a single population of molecules. The plasmins derived from Chicago I plasminogen, and also Chicago II and Chicago III plasminogens, have 100% active sites with normal amidase parameters. Basically, a single population of normal isoelectric forms were found in the Chicago III plasma, with three minor forms, about 11% of the total.The kinetic parameters have permitted us to classify the four known plasminogen variants into three different classes. The Class A homozygote (Tochigi) has both an active center defect and a charge mutation difference with normal cleavage of the Arg560-Val peptide bond; the Class B homozygote (Chicago I and Chicago II) has a Km of activation defect with impaired activator binding and normal cleavage of the Arg560-Val peptide bond; and, the Class C homozygote (Chicago III) has both a Km of activation defect with impaired activator binding and a kcat of activation defect, and impaired Arg560-Val peptide bond cleavage.

Stroke in a young adult with familial plasminogen disorder.

“…Meanwhile, streptokinase activation data shows that the abnormal plasminogen molecules not only interfere with the binding of plasminogen substrate to the activator complex, but also in the formation of the activator complex itself. The abnormality in this family is therefore most similar to the Chicago III case reported by Wohl et al 7 The most striking difference between the two reports is that Cleveland has decreased concentrations of antigen in addition to a functional defect.…”

Comparison of Human Normal, Full-Term, Fetal and Adult Plasminogen by Physical and Chemical Analyses

“…Isoelectric focusing was carried out in 5% acryl amide gels (Ortec) containing 3% bis-acrylamide, 0.2 M taurine, 0.35 M sucrose, 2% ampholines, pH 6-8 (LKB), and 0.7% ampholines, pH 3.5-10, us ing the LKB Multiphor 2117 and the LKB Power Supply 3371 E. This procedure has been described in detail [ 13], Samples of 25 pi of purified PLG at 1 mg/ ml were analyzed. Isoelectric focusing was carried out at 4 °C at 450 V for 6 h. After focusing, the gel was stained with 0.25% Coomassie brilliant blue R-250 (Bio-Rad) in 45 % methanol/10% acetic acid for 2 h at room temperature.…”

Dysplasminogenemias

“…Most of the heterozy gotes have ratios between 0.3 and 0.6 [5,[10][11][12][13][14]. Homozygotes with ratios below 0.3 [5,8], and with ratios between 0.4 and 0.9 [7,9] have been described. Pig isolated from these patient's plasmas give the same functionahantigen ratio as those determined in the plasmas.…”

“…uroki nase (u-PA) and streptokinase (SK), will usually iden tify a patient with an abnormal Pig [7,9,15]; initial velocities (vobs) in plasma are measured and com pared to initial velocities (vcaic) in a purified system with standard amounts of u-PA (250 IU) and SK (25 IU) per 0.5 ml plasma (0.1 mg pure Pig). A variant Pig will probably have lowered Pin generation rates, usually with both types of activators.…”