Molecular cloning and sequence determination of cDNAs for alpha subunits of the guanine nucleotide-binding proteins Gs, Gi, and Go from rat brain.
Molecular cloning and sequence determination of cDNAs for alpha subunits of the guanine nucleotide-binding proteins Gs, Gi, and Go from rat brain.
复制标题
大鼠脑鸟嘌呤核苷酸结合蛋白 Gs、Gi 和 Go α 亚基 cDNA 的分子克隆和序列测定。
DOI：
--
复制
发表时间：
1986
影响因子：
11.1
通讯作者：
Koichi Suzuki
中科院分区：
文献类型：
--
作者：
Hiroshi Itoh;Tohru Kozasa;Shigekazu Nagata;Shun Nakamura;Toshiaki Katada;Michio Ui;S. Iwai;E. Ohtsuka;Hiroshi Kawasaki;Koichi Suzuki
我们已经克隆了编码鸟嘌呤核苷酸结合蛋白Gs、Gi和Go的α亚基的cDNA，并测定了它们的核苷酸序列。纯化的大鼠脑GI和GOα亚基(GIα和GOα)用胰酶完全消化，并进行氨基酸序列分析。利用合成的17个氨基酸序列的探针，从大鼠C6神经胶质瘤细胞中筛选出GOα基因的编码区。然后，用GOαcDNA探针对文库进行重新筛选，以分离出几个强杂交或弱杂交克隆。从而获得了编码Giα和Gsα全序列的cDNA。通过核苷酸序列分析，推导出Gsα和Giα的氨基酸序列，分别含有394个和355个氨基酸残基(包括引发剂蛋氨酸)。计算得到Gsα和Giα的分子量分别为45,663和40,499。GOα克隆编码了一个310个氨基酸残基的序列，该序列缺少NH2末端。讨论了Gs、Gi、Go、转导蛋白和ras编码蛋白的α亚基的同源性。
We have cloned cDNAs encoding alpha subunits of the guanine nucleotide-binding proteins Gs, Gi, and Go and determined their nucleotide sequences. Purified preparations of Gi and Go alpha subunits (Gi alpha and Go alpha) from rat brain were completely digested with trypsin, and peptides were subjected to amino acid sequence analysis. By screening of a cDNA library from rat C6 glioma cells with a synthetic probe corresponding to a 17 amino acid sequence, a clone encoding the sequence of Go alpha was obtained. Then, the library was rescreened with a Go alpha cDNA probe to isolate several strongly or weakly hybridizing clones. cDNAs encoding the complete sequences of Gi alpha and Gs alpha were thus obtained. From nucleotide sequence analysis, the amino acid sequences of Gs alpha and Gi alpha were deduced; they contain 394 and 355 amino acid residues (including the initiator methionine), respectively. The calculated molecular weights for Gs alpha and Gi alpha were 45,663 and 40,499, respectively. The Go alpha clone encoded a sequence of 310 amino acid residues that lacked the NH2 terminus. The homology of the alpha subunits of Gs, Gi, Go, transducin, and ras-encoded protein is discussed.