Quantitative measurement of plasma gelsolin and its incorporation into fibrin clots

Abstract

Gelsolin is a newly recognized actin-binding protein of plasma that severs actin filaments. The concentration of plasma gelsolin was measured by three independent methods: an enzyme-linked immunosorbent assay (ELISA) and two functional assays based on the ability of gelsolin to accelerate the polymerization of actin (a "nucleating" assay) or to sever preformed actin filaments (a "cutting" assay). The gelsolin level in 56 samples of human plasma ranged between 100 and 330 micrograms/ml. The mean plasma concentrations measured by the different assays were ELISA, 207 micrograms/ml; nucleating assay, 233 micrograms/ml; cutting assay, 247 micrograms/ml. The mean serum level of gelsolin was found to be 24% lower than that of plasma when paired samples were examined (183 micrograms/ml). The difference between plasma and serum gelsolin concentrations can be accounted for by a direct interaction between gelsolin and fibrin as shown by the binding of radiolabeled gelsolin to clots made from purified fibrinogen. Further, unlabeled gelsolin inhibits the binding of the labeled species to fibrin, but hemoglobin and albumin do not. Fibrin oligomers, but not fibrinogen, alter the sedimentation characteristics of radiolabeled gelsolin in sucrose gradients. The amount of gelsolin incorporated into the clot is increased if the clots are made in the presence of actin filaments or fibronectin. Thus, serum levels of gelsolin are lower than plasma levels because of an interaction between gelsolin and fibrin clots.