Lipkin V M, Mirzoeva S F, Dranitsyna S M, Moshniakov M V, Petrov V M
Bioorg Khim. 1989 Jan;15(1):5-17.
The catalytic component of calmodulin-independent adenylate cyclase of cattle cerebral cortex was solubilized and purified to the homogeneous state. The conditions for preparative obtaining of the enzyme on the column with immobilized antibodies to adenylate cyclase were found. These antibodies were proved to interact with the calmodulin-independent rather than the calmodulin-dependent form of the enzyme. Molecular mass of the calmodulin-independent adenylate cyclase determined electrophoretically is 140 +/- 10 kDa. Amino acid composition of the enzyme and sequences of its fragments (in total 300 amino acid residues) obtained upon treatment with lysyl-specific proteinase from Achromobacter liticus were determined. Clone containing a cDNA 605 bp insertion coding for the 183 amino acid residue fragment of adenylate cyclase was isolated from the bovine brain cDNA library. Homology of this fragment to the known sequences of Escherichia coli and Bordetella pertussis adenylate cyclases was revealed.
牛大脑皮层中不依赖钙调蛋白的腺苷酸环化酶的催化成分被溶解并纯化至均一状态。发现了在固定有腺苷酸环化酶抗体的柱上制备该酶的条件。已证明这些抗体与不依赖钙调蛋白而非依赖钙调蛋白形式的酶相互作用。通过电泳测定，不依赖钙调蛋白的腺苷酸环化酶的分子量为140±10 kDa。测定了该酶的氨基酸组成及其在用来自浅青紫链霉菌的赖氨酰特异性蛋白酶处理后获得的片段序列（总共300个氨基酸残基）。从牛脑cDNA文库中分离出一个克隆，其包含编码腺苷酸环化酶183个氨基酸残基片段的605 bp cDNA插入片段。揭示了该片段与大肠杆菌和百日咳博德特氏菌腺苷酸环化酶已知序列的同源性。
