Identification of gelsolin, a Ca2+-dependent regulatory protein of actin gel-sol transformation, and its intracellular distribution in a variety of cells and tissues

Abstract

Antiserum prepared against gelsolin, a major Ca2+-dependent regulatory protein of actin gel-sol transformation in rabbit lung macrophages, was used to detect the presence of proteins immunologically related to gelsolin in a variety of cells and tissues. Cell extracts were electrophoresed on polyacrylamide gels, and replicas of the gels on cellulose nitrate paper were stained by an indirect immunohistochemical technique. A single band of crossreactive material which comigrates with macrophage gelsolin is found in at least nine different kinds of cells and tissues derived from rabbits and humans and in four lines of cultured cells from humans and rats. Gelsolin was also identified in human serum and plasma, raising the possibility that it may contribute to the clearance of actin from the circulatory system. Using this antiserum, we demonstrated, by indirect immunofluorescent staining of acetone-fixed macrophages and polymorphonuclear leukocytes, that gelsolin resides in the cortical cytoplasm and that during phagocytosis it is concentrated in pseudopodia engulfing particles to be ingested, an area of the cytoplasm actively engaged in movement. In longitudinal cryostat sections of contracted rabbit skeletal muscle, antigelsolin staining was associated with the I-band of the myofibril, suggesting that it may be involved, by an as yet undefined mechanism, in skeletal muscle function. In rabbit intestinal epithelial cells, gelsolin was associated with the cytoplasm and the terminal web region of the brush border, a localization distinct from that previously reported for villin, a structurally and functionally similar protein isolated from the brush borders of chicken intestinal epithelial cells. In conclusion, our findings support the idea that gelsolin is involved in the regulation of movement and suggest that gelsolin-mediated Ca2+-regulation of actin cytoskeletal structure, first characterized in macrophages, may be of general importance.