A functional assay for proteins involved in establishing an epithelial occluding barrier: identification of a uvomorulin-like polypeptide.
A functional assay for proteins involved in establishing an epithelial occluding barrier: identification of a uvomorulin-like polypeptide.
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涉及建立上皮遮挡屏障的蛋白质的功能分析：识别紫外线蛋白样多肽。
DOI：
10.1083/jcb.102.2.457
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发表时间：
1986-02
影响因子：
7.8
通讯作者：
SIMONS, K
中科院分区：
文献类型：
--
作者：
GUMBINER, B;SIMONS, K
已经开发了一种功能测定法来鉴定参与上皮紧密连接形成的细胞表面蛋白。跨上皮电阻用于测量在硝酸纤维素滤膜上培养的Madin-Darby犬肾（MDCK）细胞单层中完整紧密连接的存在。使用的菌株I MDCK细胞具有大于2，000欧姆的跨单层电阻。cm2.当单层细胞在37 ℃下无Ca 2+孵育时，细胞间连接打开，跨单层电阻下降到裸滤器的值，即，小于40欧姆。cm2.当Ca 2+恢复时，细胞连接重新封闭，电阻迅速恢复。多克隆抗体对完整的MDCK细胞抑制钙依赖的电阻恢复时，施加到单层已开放的钙去除。不需要细胞表面分子的交联，因为单价Fab'片段也被抑制。相反，识别MDCK细胞表面特异性蛋白质的各种其他抗体未能抑制抗性的恢复。单克隆抗体已被提出并筛选其抑制抗性恢复的能力。已经获得了一种这样的单克隆抗体，其染色MDCK细胞的侧表面。该抗体，rr 1，识别MDCK细胞提取物中的118- kD多肽和在Ca 2+存在下通过胰蛋白酶消化从细胞表面释放的81-kD片段。顺序免疫沉淀抗体rr 1和一个单克隆抗体，以uvomorulin表明，这种多肽与uvomorulin。的作用，uvomorulin样和肝细胞粘附分子（L-CAM）样多肽在上皮闭塞屏障的建立进行了讨论。
A functional assay has been developed to identify cell surface proteins involved in the formation of epithelial tight junctions. Transepithelial electrical resistance was used to measure the presence of intact tight junctions in monolayers of Madin-Darby canine kidney (MDCK) cells cultured on nitrocellulose filters. The strain I MDCK cells used have a transmonolayer resistance greater than 2,000 ohm . cm2. When the monolayers were incubated at 37 degrees C without Ca2+, the intercellular junctions opened and the transmonolayer resistance dropped to the value of a bare filter, i.e., less than 40 ohm . cm2. When Ca2+ was restored, the cell junctions resealed and the resistance recovered rapidly. Polyclonal antibodies raised against intact MDCK cells inhibited the Ca2+-dependent recovery of electrical resistance when applied to monolayers that had been opened by Ca2+ removal. Cross- linking of cell surface molecules was not required because monovalent Fab' fragments also inhibited. In contrast, a variety of other antibodies that recognize specific proteins on the MDCK cell surface failed to inhibit the recovery of resistance. Monoclonal antibodies have been raised and screened for their ability to inhibit resistance recovery. One such monoclonal antibody has been obtained that stained the lateral surface of MDCK cells. This antibody, rr1, recognized a 118- kD polypeptide in MDCK cell extracts and an 81-kD fragment released from the cell surface by trypsinization in the presence of Ca2+. Sequential immunoprecipitation with antibody rr1 and a monoclonal antibody to uvomorulin showed that this polypeptide is related to uvomorulin. The role of uvomorulin-like and liver cell adhesion molecule (L-CAM)-like polypeptides in the establishment of the epithelial occluding barrier is discussed.
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影响因子：
64.5
作者：
DAMSKY, CH;RICHA, J;BUCK, CA
通讯作者：
BUCK, CA
DOI：
10.1083/jcb.101.4.1307
发表时间：
1985-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Behrens J;Birchmeier W;Goodman SL;Imhof BA
通讯作者：
Imhof BA
DOI：
10.1083/jcb.100.1.327
发表时间：
1985-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Boller K;Vestweber D;Kemler R
通讯作者：
Kemler R
影响因子：
2.4
作者：
CLAUDE, P
通讯作者：
CLAUDE, P
DOI：
10.1083/jcb.96.1.230
发表时间：
1983-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hubbard AL;Ma A
通讯作者：
Ma A