title: Studies of C1 inactivator-plasma kallikrein complexes in purified systems and in plasma.
abstract: An enzyme-linked immunosorbent assay (ELISA) has been developed for the quantification of C1 inactivator-kallikrein (C1In-K) complexes. The formation of complexes assayed by this method parallelled the inhibition of plasma kallikrein esterase activity by C1 inactivator in purified systems. C1In-K complexes were detected when a final concentration of 5.7 nM plasma kallikrein was added to plasma, equivalent to the activation of 1% of the plasma prekallikrein. Exogenous Hageman factor fragment added to plasma induced the rapid formation of C1In-K complexes, whereas there was an appreciable delay when the plasma contact system was activated by the addition of kaolin. In both systems, the rate of formation and final amount of complex generated were directly related to the concentration of Hageman factor fragment or of kaolin added, indicating that this proteolytic pathway is tightly regulated. C1In-K complexes were not generated by kaolin in plasma congenitally deficient in Hageman factor or prekallikrein or by kallikrein in hereditary angioedema plasma deficient in C1 inactivator, thus confirming the specificity of the assay. Sucrose gradient ultracentrifugation studies showed plasma C1In-K complexes to have a molecular weight consistent with a 1:1 molar complex. In contrast, the complex displayed an anomalously high molecular weight on gel filtration chromatography. These data demonstrate that a sensitive and specific probe has been developed for documenting plasma kallikrein activation.
mesh_terms: D000799:Angioedema / Q000097:blood; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003174:Complement C1 Inactivator Proteins / Q000097:blood*; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005174:Factor XII / Q000494:pharmacology; D006801:Humans; D007124:Immunoenzyme Techniques; D007610:Kallikreins / Q000037:antagonists & inhibitors / Q000097:blood*; D007616:Kaolin / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D011288:Prekallikrein / Q000502:physiology
chemical_list: D003174:Complement C1 Inactivator Proteins; D010446:Peptide Fragments; D007616:Kaolin; D005174:Factor XII; D011288:Prekallikrein; D007610:Kallikreins