Molecular structural studies of gap junction protein compositions.

间隙连接蛋白组合物的分子结构研究。

基本信息

批准号：
05454136
负责人：
SHIBATA Yosaburo
金额：
$ 3.46万
依托单位：
KYUSYU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

Gap junction proteins of connexin (Cx) family were examined by using specific rabbit antibodies raised against type-specific peptide sequence domains at the single connexon channel level and on the tissue distiribution and cell type secificity.1. Single gap junction channel (connexon) particles isolated from rat and guinea pig liver by mild detergent extraction showed doughnut-shaped unit particles with central depressions in negative staining and low angle rotary shadowing electron microscopy. Further purification by high performance liquid chromatography followed by affinity chromatogram on either anti Cx26 of anti-Cx32 specific antibody column yielded single western blot pattern without appreciable cosedimentations of other connexins. This result may indicate that single connexon hexamers are composed of either Cx26 of Cx32 only.2. In contrast to the salivary glands which contained both Cx32 and Cx26, lipid secreting Harderian glands showed only Cx26 immunofluorescence in the acinar cells. Myoepithelial cells showed Cx43. In mammary glands, Cx26 and Cx32 in acinar cells and Cx43 in myoepithelial cells revealed dynamic changes of expression during pregnancy, parturition, lactation and weaning period at RNA transcription, protein production, and phosphorylation states, indicating the activity-related expression of gap junction proteins thus consituting functionally distinct cell groups.These results indicate that differential expressions of various gap junction proteins are closely correlated with cell type specific functions and activities.

通过使用针对单个连接频道水平的类型特异性肽序列域的特定兔抗体以及在组织膨胀和细胞类型的确定性上，使用针对特异性特异性肽序列结构域提出的特定兔抗体检查了连接蛋白（CX）家族的间隙连接蛋白。1。通过轻度清洁剂提取从大鼠和豚鼠肝分离的单间隙连接通道（连接）颗粒显示出具有负染色和低角度旋转阴影电子显微镜的中心降低的坚果形单位颗粒。高性能液相色谱进一步纯化，然后在抗CX32特异性抗体柱上的任何一种抗CX26上进行亲和力色谱图产生了单个蛋白质印迹模式，而没有其他连接素的可观同谋。该结果可能表明单个连接六聚体仅由CX32的CX26组成2。与含有CX32和CX26的唾液腺相反，脂质分泌硬腺仅显示腺泡细胞中的CX26免疫荧光。肌上皮细胞显示CX43。 In mammary glands, Cx26 and Cx32 in acinar cells and Cx43 in myoepithelial cells revealed dynamic changes of expression during pregnancy, parturition, lactation and weaning period at RNA transcription, protein production, and phosphorylation states, indicating the activity-related expression of gap junction proteins thus consituting functionally distinct cell groups.These results indicate that differential expressions of various gap连接蛋白与细胞类型特异性功能和活性密切相关。