Sampath R, Gallagher P J, Pavalko F M
Department of Physiology and Biophysics, Indiana University School of Medicine, Indianapolis, Indiana 46202-5120, USA.
J Biol Chem. 1998 Dec 11;273(50):33588-94. doi: 10.1074/jbc.273.50.33588.
Circulating leukocytes are nonadherent but bind tightly to endothelial cells following activation. The increased avidity of leukocyte integrins for endothelial ligands following activation is regulated, in part, by interaction of the beta2 subunit cytoplasmic tail with the actin cytoskeleton. We propose a mechanism to explain how tethering of the actin cytoskeleton to leukocyte integrins is regulated. In resting leukocytes, beta2 integrins are constitutively linked to the actin cytoskeleton via the protein talin. Activation of cells induces proteolysis of talin and dissociation from the beta2 tail. This phase is transient, however, and is followed by reattachment of actin filaments to integrins that is mediated by the protein alpha-actinin. The association of alpha-actinin with integrins may stabilize the cytoskeleton and promote firm adhesion to and migration across the endothelium. Glutathione S-transferase-beta2 tail fusion protein/mutagenesis experiments suggest that the affinity of alpha-actinin binding to the beta2 tail is regulated by a change in the conformation of the tail that unmasks a cryptic alpha-actinin binding domain. Positive and inhibitory domains within the beta2 tail regulate alpha-actinin binding: a single 11-amino acid region (residues 736-746) is necessary and sufficient for alpha-actinin binding, and a regulatory domain between residues 748-762 inhibits constitutive association of the beta2 tail with alpha-actinin.
循环白细胞是非黏附性的,但激活后会紧密结合到内皮细胞上。激活后白细胞整合素对内皮配体的亲和力增加,部分是由β2亚基胞质尾与肌动蛋白细胞骨架的相互作用调节的。我们提出一种机制来解释肌动蛋白细胞骨架与白细胞整合素的连接是如何被调节的。在静息白细胞中,β2整合素通过踝蛋白与肌动蛋白细胞骨架组成性连接。细胞激活诱导踝蛋白的蛋白水解并从β2尾解离。然而,这个阶段是短暂的,随后是由α辅肌动蛋白介导的肌动蛋白丝重新附着到整合素上。α辅肌动蛋白与整合素的结合可能稳定细胞骨架并促进与内皮的牢固黏附以及跨内皮迁移。谷胱甘肽S-转移酶-β2尾融合蛋白/诱变实验表明,α辅肌动蛋白与β2尾的结合亲和力是由尾构象的变化调节的,这种变化会暴露一个隐蔽的α辅肌动蛋白结合结构域。β2尾内的正向和抑制结构域调节α辅肌动蛋白的结合:一个单一的11个氨基酸区域(第736 - 746位残基)对于α辅肌动蛋白结合是必需且足够的,并且第748 - 762位残基之间的调节结构域抑制β2尾与α辅肌动蛋白的组成性结合。