PERTUSSIS TOXIN SUBSTRATE, THE PUTATIVE NI COMPONENT OF ADENYLYL CYCLASES, IS AN ALPHA-BETA-HETERODIMER REGULATED BY GUANINE-NUCLEOTIDE AND MAGNESIUM
PERTUSSIS TOXIN SUBSTRATE, THE PUTATIVE NI COMPONENT OF ADENYLYL CYCLASES, IS AN ALPHA-BETA-HETERODIMER REGULATED BY GUANINE-NUCLEOTIDE AND MAGNESIUM
复制标题
DOI：
10.1073/pnas.80.14.4276
复制
发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
MANCLARK, CR
中科院分区：
文献类型：
--
作者：
CODINA, J;HILDEBRANDT, J;MANCLARK, CR
从人红细胞膜纯化腺苷酸环化酶[腺苷酸环化酶; ATP焦磷酸-裂解酶（环化），EC 4.6.1.1]的鸟嘌呤核苷酸-和Mg 2 +-结合刺激调节组分（Ns）的方案中的最后一步涉及在羟基磷灰石（HAP）上的色谱法，其产生2个级分。第1级分（HAP I）主要含有2种肽，在十二烷基硫酸钠/聚丙烯酰胺凝胶电泳时，迁移的MW值为39，000和35，000。第二部分（HAP II）主要含有由MW 42，000和35，000的2种肽形成的Ns。HAP I，MW 39，000肽显示为百日咳博德特氏菌的ADP-核糖基化毒素（百日咳毒素）的底物。在蔗糖密度梯度离心后，HAP I的MW 39，000和MW 35，000肽均在约100 ℃迁移。4 S。在离心之前用鸟嘌呤核苷酸和Mg 2+处理HAP I导致两种肽向2S迁移的协调变化。假设HAP I含有α。β的异二聚体蛋白质，其由αMW 39，000的亚基和β亚基。MW 35，000的亚基。该蛋白质在鸟嘌呤核苷酸和Mg 2+的影响下解离成其单独的α。和β亚单位。因为先前的研究已经表明，用百日咳毒素处理细胞和细胞膜导致抑制腺苷酸环化酶活性的激素的作用减弱，并且因为这种作用与MW 40，000肽的ADP-核糖基化相关。腺苷酸环化酶的一种鸟嘌呤核苷酸和Mg 2+结合抑制调节成分，即，倪，被认为是被净化了。
The final step in a scheme for the purification of the guanine nucleotide- and Mg2+-binding stimulatory regulatory component (Ns) of adenylyl cyclase [adenylate cyclase; ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] from human erythrocyte membranes involves chromatography over hydroxylapatite (HAP) which yields 2 fractions. The 1st fraction (HAP I) contains predominantly 2 peptides that, upon sodium dodecyl sulfate/polyacrylamide gel electrophoresis, migrate with MW values of 39,000 and 35,000. The 2nd fraction (HAP II) contains predominantly Ns formed of 2 peptides of MW 42,000 and 35,000. The HAP I, MW 39,000 peptide is shown to be a substrate for the ADP-ribosylating toxin of Bordetella pertussis (pertussis toxin). Upon sucrose density gradient centrifugation, both the MW 39,000 and the MW 35,000 peptides of HAP I migrate at .apprx. 4 S. Treatment of HAP I with guanine nucleotide and Mg2+ prior to centrifugation results in a coordinated change in the migration of both peptides to 2 S. It is postulated that HAP I contains an .alpha..beta. heterodimeric protein composed of an .alpha. subunit of MW 39,000 and a .beta. subunit of MW 35,000. This protein dissociates under the influence of guanine nucleotides and Mg2+ into its individual .alpha. and .beta. subunits. Because previous studies have shown that treatment of cells and cell membranes with pertussis toxin results in attenuation of the effects of hormones that inhibit adenylyl cyclase activity, and because this effect correlates with the ADP-ribosylation of a MW 40,000 peptide. A guanine nucleotide- and Mg2+-binding inhibitiory regulatory component of adenylyl cyclases i.e., the Ni, is thought to have been purified.