Eisenschlos C, Flawiá M M, Torruella M, Torres H N
Biochem J. 1986 May 15;236(1):185-91. doi: 10.1042/bj2360185.
Trypanosoma cruzi adenylate cyclase catalytic subunits may interact with regulatory factors from rat liver membranes, reconstituting heterologous systems which are catalytically active in assay mixtures containing MgATP. The systems show stimulatory responses to glucagon and guanosine 5'-[beta gamma-imido]triphosphate (p[NH]ppG) or fluoride. Reconstitution was obtained by three different methods: fusion of rat liver membranes (pretreated with N-ethylmaleimide) to T. cruzi membranes; interaction of detergent extracts of rat liver membranes with T. cruzi membranes; or interaction of purified preparations of T. cruzi adenylate cyclase and of liver membrane factors in phospholipid vesicles. The liver factors responsible for the guanine nucleotide effect were characterized as the NS protein. Data also indicate that reconstitution requires the presence of a membrane substrate.
克氏锥虫腺苷酸环化酶催化亚基可能与大鼠肝细胞膜的调节因子相互作用,重建在含有MgATP的测定混合物中具有催化活性的异源系统。这些系统对胰高血糖素、鸟苷5'-[βγ-亚氨基]三磷酸(p[NH]ppG)或氟化物表现出刺激反应。通过三种不同方法实现了重建:将大鼠肝细胞膜(用N-乙基马来酰亚胺预处理)与克氏锥虫膜融合;大鼠肝细胞膜的去污剂提取物与克氏锥虫膜相互作用;或克氏锥虫腺苷酸环化酶的纯化制剂与肝膜因子在磷脂囊泡中的相互作用。负责鸟嘌呤核苷酸效应的肝因子被鉴定为NS蛋白。数据还表明重建需要膜底物的存在。