Werner R, Levine E, Rabadan-Diehl C, Dahl G
Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Florida 33101.
Proc Biol Sci. 1991 Jan 22;243(1306):5-11. doi: 10.1098/rspb.1991.0002.
Carboxyl-terminal deletion mutants of the gap junction protein connexin32 were tested in the oocyte cell-cell channel assay. Oocytes expressing a mutant lacking 58 carboxyl terminal amino acids were found to exhibit junctional conductances of the same magnitude as oocytes expressing wild-type connexin32. The gating properties of the channels formed by this mutant of connexin32 with respect to transjunctional voltage and cytoplasmic acidification are indistinguishable from those found with wild-type connexin32 channels. This includes a novel pH-dependent voltage gate. In another mutant, two carboxyl terminal serine residues, Ser233 and Ser240, were replaced by Asn residues. This double mutant has properties indistinguishable from wild-type connexin32, suggesting that phosphorylation of either of these serines is not required for channel opening.
在卵母细胞细胞间通道检测中对间隙连接蛋白连接蛋白32的羧基末端缺失突变体进行了测试。发现表达缺失58个羧基末端氨基酸的突变体的卵母细胞,其连接电导与表达野生型连接蛋白32的卵母细胞相同。该连接蛋白32突变体形成的通道在跨连接电压和细胞质酸化方面的门控特性与野生型连接蛋白32通道的门控特性没有区别。这包括一种新的pH依赖性电压门。在另一个突变体中,两个羧基末端丝氨酸残基Ser233和Ser240被Asn残基取代。这个双突变体的特性与野生型连接蛋白32没有区别,这表明这些丝氨酸中的任何一个的磷酸化对于通道开放不是必需的。