Expression of VAMP-2-like protein in kidney collecting duct intracellular vesicles. Colocalization with Aquaporin-2 water channels.
Expression of VAMP-2-like protein in kidney collecting duct intracellular vesicles. Colocalization with Aquaporin-2 water channels.
复制标题
肾集合管细胞内囊泡中VAMP-2样蛋白的表达。
DOI：
--
复制
发表时间：
1995
影响因子：
15.9
通讯作者：
M. Knepper
中科院分区：
文献类型：
--
作者：
S. Nielsen;D. Marples;H. Birn;M. Mohtashami;N. O. Dalby;M. Trimble;M. Knepper
体内水分平衡由加压素控制，加压素通过胞内囊泡和质膜之间的囊泡运输来调节肾集合管细胞中的水通道蛋白-2（AQP 2）水通道。为了研究集合管细胞中AQP 2的囊泡运输和加压素调节所涉及的分子装置，我们测试了在突触囊泡中表达的靶向蛋白，即囊泡相关膜蛋白1和2（VAMP 1和2），是否在肾脏集合管中表达。免疫印迹显示特异性标记的VAMP 2（18-kD带），但不VAMP 1从肾内髓质制备的膜组分。使用预吸附抗体或免疫前血清的对照为阴性。在脑膜囊泡和纯化的突触囊泡的免疫印迹中检测到相同分子大小的条带。肾膜中的VAMP 2被破伤风毒素切割，揭示了破伤风毒素敏感的VAMP同源物。类似地，破伤风毒素在突触囊泡中切割VAMP 2。在肾内髓质中，VAMP 2主要表达于富含细胞内囊泡的膜组分中，在富含质膜的组分中很少或没有VAMP 2。这是证实了免疫细胞化学使用半薄冷冻切片，主要是囊泡标记的集合管主细胞，没有标记的闰细胞。VAMP 2免疫标记与AQP 2标记共定位于胞内囊泡中，如在分离的囊泡的双重免疫标记后通过免疫电子显微镜测定的。对1，310个囊泡的定量分析揭示了同一囊泡中AQP 2和VAMP 2的高度显著相关性（P < 0.0001）。此外，通过免疫印迹证实了用抗VAMP 2抗体免疫分离的囊泡中存在AQP 2。总之，VAMP 2，神经元SNARE复合物的一个组成部分，在携带AQP 2的囊泡中表达，表明在加压素调节的囊泡运输AQP 2水通道中的作用。
Body water balance is controlled by vasopressin, which regulates Aquaporin-2 (AQP2) water channels in kidney collecting duct cells by vesicular trafficking between intracellular vesicles and the plasma membrane. To examine the molecular apparatus involved in vesicle trafficking and vasopressin regulation of AQP2 in collecting duct cells, we tested if targeting proteins expressed in the synaptic vesicles, namely vesicle-associated membrane proteins 1 and 2 (VAMP1 and 2), are expressed in kidney collecting duct. Immunoblotting revealed specific labeling of VAMP2 (18-kD band) but not VAMP1 in membrane fractions prepared from kidney inner medulla. Controls using preadsorbed antibody or preimmune serum were negative. Bands of identical molecular size were detected in immunoblots of brain membrane vesicles and purified synaptic vesicles. VAMP2 in kidney membranes was cleaved by tetanus toxin, revealing a tetanus toxin-sensitive VAMP homologue. Similarly, tetanus toxin cleaved VAMP2 in synaptic vesicles. In kidney inner medulla, VAMP2 was predominantly expressed in the membrane fraction enriched for intracellular vesicles, with little or no VAMP2 in the plasma membrane enriched fraction. This was confirmed by immunocytochemistry using semithin cryosections, which showed mainly vesicular labeling in collecting duct principal cells, with no labeling of intercalated cells. VAMP2 immunolabeling colocalized with AQP2 labeling in intracellular vesicles, as determined by immunoelectron microscopy after double immunolabeling of isolated vesicles. Quantitative analysis of 1,310 vesicles revealed a highly significant association of both AQP2 and VAMP2 in the same vesicles (P < 0.0001). Furthermore, the presence of AQP2 in vesicles immunoisolated with anti-VAMP2 antibodies was confirmed by immunoblotting. In conclusion, VAMP2, a component of the neuronal SNARE complex, is expressed in vesicles carrying AQP2, suggesting a role in vasopressin-regulated vesicle trafficking of AQP2 water channels.
登录
查看更多内容
DOI：
10.1016/s0021-9258(19)49748-2
发表时间：
1992-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Cynthia A. Corley;Cain;William S Trimblez;Gustav E Lienhardq
通讯作者：
Cynthia A. Corley;Cain;William S Trimblez;Gustav E Lienhardq
DOI：
10.1172/jci116798
发表时间：
1993-10
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Barbara L. Smith;Ruben Baumgarten;Søren Nielsen;D. M. Raben;M. Zeidel;P. Agre
通讯作者：
Barbara L. Smith;Ruben Baumgarten;Søren Nielsen;D. M. Raben;M. Zeidel;P. Agre
DOI：
10.1073/pnas.90.15.7275
发表时间：
1993-08-01
影响因子：
11.1
作者：
NIELSEN, S;SMITH, BL;AGRE, P
通讯作者：
AGRE, P
DOI：
10.1073/pnas.90.24.11663
发表时间：
1993-12-15
影响因子：
11.1
作者：
NIELSEN, S;DIGIOVANNI, SR;HARRIS, HW
通讯作者：
HARRIS, HW