Shlatz L J, Schwartz I L, Kinne-Saffran E, Kinne R
J Membr Biol. 1975 Nov 7;24(2):131-44. doi: 10.1007/BF01868619.
Free flow electrophoresis was employed to separate renal cortical plasma membranes into luminal (brush border microvilli) and contraluminal (basal-lateral membrane) fractions. During the separation adenylate cyclase activity was found to parallel the activity of Na+-K+-activated ATPase, an enzyme which is present in contraluminal but not in luminal membranes. In the basal-lateral membrane fraction the specific activities of adenylate cyclase and Na+-K+-activated ATPase were 4.4 and 4.6 times greater, respectively, than in the brush border fraction. The adenylate cyclase of the basal-lateral membrane fraction was specifically stimulated by parathyroid hormone which maximally increased enzyme activity eightfold. The biologically active (1-34) peptide fragment of paratyhroid hormone produced a 350% increase in adenylate cyclase activity. In contrast, calcitonin, epinephrine and vasopressin maximally stimulated the enzyme by only 55, 35 and 30%, respectively. These results indicate that adenylate cyclase, specifically stimulated by parathyroid hormone, is distributed preferentially in the contraluminal region of the plasma membrane of renal cortical epithelial cells.
采用自由流动电泳法将肾皮质质膜分离为管腔面(刷状缘微绒毛)和非管腔面(基底外侧膜)组分。在分离过程中,发现腺苷酸环化酶活性与钠钾激活的ATP酶活性平行,该酶存在于非管腔面而不存在于管腔面膜中。在基底外侧膜组分中,腺苷酸环化酶和钠钾激活的ATP酶的比活性分别比刷状缘组分高4.4倍和4.6倍。基底外侧膜组分的腺苷酸环化酶受到甲状旁腺激素的特异性刺激,酶活性最大增加了8倍。甲状旁腺激素的生物活性(1-34)肽片段使腺苷酸环化酶活性增加了350%。相比之下,降钙素、肾上腺素和血管加压素分别仅使该酶的活性最大刺激55%、35%和30%。这些结果表明,受甲状旁腺激素特异性刺激的腺苷酸环化酶优先分布于肾皮质上皮细胞质膜的非管腔区域。