Identification of gelsolin, a Ca2+-dependent regulatory protein of actin gel-sol transformation, and its intracellular distribution in a variety of cells and tissues.
Identification of gelsolin, a Ca2+-dependent regulatory protein of actin gel-sol transformation, and its intracellular distribution in a variety of cells and tissues.
复制标题
DOI：
10.1083/jcb.91.3.901
复制
发表时间：
1981-12
期刊：
影响因子：
--
通讯作者：
Fattoum A
中科院分区：
文献类型：
--
作者：
Yin HL;Albrecht JH;Fattoum A
gelsolin是兔肺巨噬细胞肌动蛋白凝胶-溶胶转化的主要Ca2+依赖性调节蛋白，制备了针对gelsolin的抗血清，用于检测多种细胞和组织中与gelsolin免疫相关的蛋白的存在。细胞提取物在聚丙烯酰胺凝胶上电泳，复制的凝胶在硝酸纤维素纸上用间接免疫组织化学技术染色。至少在兔和人类的九种不同的细胞和组织以及人类和大鼠的四种培养细胞中发现了与巨噬细胞凝胶相结合的单条带交叉反应物质。明胶也在人血清和血浆中被发现，这提高了它可能有助于清除循环系统中的肌动蛋白的可能性。使用这种抗血清，我们通过对丙酮固定巨噬细胞和多形核白细胞的间接免疫荧光染色证明，凝胶蛋白存在于皮质细胞质中，在吞噬过程中，凝胶蛋白集中在吞噬颗粒的伪足中，这是细胞质中积极参与运动的区域。在兔骨骼肌收缩的纵向低温切片中，抗凝胶蛋白染色与肌原纤维的i带相关，表明它可能以一种尚未确定的机制参与骨骼肌功能。在兔肠上皮细胞中，凝胶蛋白与细胞质和刷状边缘的终网区域相关，这与先前报道的绒毛蛋白（一种从鸡肠上皮细胞刷状边缘分离出来的结构和功能相似的蛋白）的定位不同。总之，我们的研究结果支持gelsolin参与运动调节的观点，并表明gelsolin介导的Ca2+调节肌动蛋白细胞骨架结构（首先在巨噬细胞中表征）可能具有普遍重要性。
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.