Le Vine H, Cuatrecasas P
Biochim Biophys Acta. 1981 Feb 5;672(3):248-61. doi: 10.1016/0304-4165(81)90291-9.
A cytosolic, macromolecular factor required for the cholera toxin-dependent activation of pigeon erythrocyte adenylate cyclase and cholera toxin-dependent ADP-ribosylation of a membrane-bound 43,000 dalton polypeptide has been purified 1100-fold from horse erythrocyte cytosol using organic solvent precipitation and heat treatment. This factor, 13,000 daltons, does not absorb to anionic or cationic exchange resins, is sensitive to trypsin or 10% trichloroacetic acid and is not extractable by diethyl ether. Activation of adenylate cyclase by cholera toxin requires the simultaneous presence of ATP (including possible trace GTP), NAD+, dithiothreitol, cholera toxin, membranes and the cytosolic macromolecular factor. Reversal of cholera toxin activation of adenylate cyclase, and of the toxin-dependent ADP-ribosylation, requires the presence of the cytosolic factor. The ability of the purified cytosolic factor to influence the hormonal sensitivity of liver membrane adenylate cyclase may provide clues to its physiological functions.
一种胞质大分子因子已从马红细胞胞质溶胶中通过有机溶剂沉淀和热处理纯化了1100倍，它是霍乱毒素依赖的鸽红细胞腺苷酸环化酶激活以及膜结合的43,000道尔顿多肽的霍乱毒素依赖的ADP核糖基化所必需的。这种13,000道尔顿的因子不吸附于阴离子或阳离子交换树脂，对胰蛋白酶或10%三氯乙酸敏感，且不能被乙醚提取。霍乱毒素激活腺苷酸环化酶需要同时存在ATP（包括可能的微量GTP）、NAD⁺、二硫苏糖醇、霍乱毒素、膜和胞质大分子因子。霍乱毒素激活腺苷酸环化酶以及毒素依赖的ADP核糖基化的逆转需要胞质因子的存在。纯化的胞质因子影响肝膜腺苷酸环化酶激素敏感性的能力可能为其生理功能提供线索。
