Ross E M, Gilman A G
J Biol Chem. 1977 Oct 25;252(20):6966-9.
Adenylate cyclase activity in a Lubrol 12A9 extract of wild type S49 lymphoma plasma membranes is completely inactivated by incubation at 37 degrees for 20 min. Activity is restored by mixing this heated extract of wild type membranes with an unheated detergent extract of membranes from a variant clone that lacks measureable adenylate cyclase activity (AC-). The factor(s) donated by the AC- extract is labile to heating at 30 degrees (t1/2 = 3 min) or to treatment with N-ethylmaleimide or trypsin. The factor(s) donated by the heated wild type extract is also sensitive to proteases or N-ethylmaleimide. This extract displays more complex inactivation kinetics at 50 degrees, consistent with the existence of separate factors necessary for the stimulatory effects of NaF and guanyl-5'-yl-imidodiphosphate. We suggest that at least two proteins are necessary for adenylate cyclase activity and that one of these is retained in the phenotypically adenylate cyclase-deficient variant.
野生型S49淋巴瘤质膜的Lubrol 12A9提取物中的腺苷酸环化酶活性，在37℃孵育20分钟后会完全失活。将这种加热过的野生型膜提取物与来自一个缺乏可测量腺苷酸环化酶活性的变异克隆（AC-）的膜的未加热去污剂提取物混合，活性得以恢复。AC-提取物提供的因子对30℃加热（半衰期 = 3分钟）、N-乙基马来酰亚胺处理或胰蛋白酶处理不稳定。加热过的野生型提取物提供的因子对蛋白酶或N-乙基马来酰亚胺也敏感。该提取物在50℃时表现出更复杂的失活动力学，这与氟化钠和鸟苷-5'-基-亚氨基二磷酸刺激作用所需的不同因子的存在一致。我们认为，腺苷酸环化酶活性至少需要两种蛋白质，其中一种保留在表型上缺乏腺苷酸环化酶的变异体中。
