Kaetzel M A, Chan H C, Dubinsky W P, Dedman J R, Nelson D J
Department of Physiology and Biophysics, University of Cincinnati College of Medicine, Ohio 45267-0576.
J Biol Chem. 1994 Feb 18;269(7):5297-302.
The cellular function of annexin IV was evaluated by correlating tissue expression, cellular localization, and whole-cell electrophysiology. Immunolocalization and biochemical data demonstrate that annexin IV is concentrated along the apical membranes of many epithelia. Introduction of purified exogenous annexin IV into colonic T84 cells through a patch pipette specifically prevented Ca(2+)-dependent Cl- current activation. Affinity-purified antibody against annexin IV applied in the same manner enhanced the activation. Reduction of the endogenous level of annexin IV with a derivatized oligodeoxynucleotide antisense to annexin IV mRNA lowered the threshold for the Ca(2+)-induced current response, mimicking the enhancement of current activation exerted by anti-annexin IV antibody. The inhibitory effect of annexin IV on Ca(2+)-dependent Cl- conductance represents a novel mechanism by which Ca(2+)-binding proteins modulate membrane channel activity.
通过关联组织表达、细胞定位和全细胞电生理学来评估膜联蛋白IV的细胞功能。免疫定位和生化数据表明,膜联蛋白IV集中在许多上皮细胞的顶端膜上。通过膜片移液管将纯化的外源性膜联蛋白IV引入结肠T84细胞中,特异性地阻止了Ca(2+)依赖性Cl-电流激活。以相同方式应用的针对膜联蛋白IV的亲和纯化抗体增强了激活作用。用与膜联蛋白IV mRNA反义的衍生化寡脱氧核苷酸降低膜联蛋白IV的内源性水平,降低了Ca(2+)诱导电流反应的阈值,模拟了抗膜联蛋白IV抗体对电流激活的增强作用。膜联蛋白IV对Ca(2+)依赖性Cl-电导的抑制作用代表了一种新机制,即Ca(2+)结合蛋白调节膜通道活性。