Pozdnyakov N, Margulis A, Sitaramayya A
Eye Research Institute, Oakland University, Rochester, Michigan 48309, USA.
Biochem Biophys Res Commun. 1997 Jun 27;235(3):482-6. doi: 10.1006/bbrc.1997.6811.
Five ciliary body membrane proteins were labeled when incubated with (adenylate-32P)NAD. Nitric oxide donors stimulated the labeling of 64, 40, and 29-30 kDa proteins and inhibited that of 58 and 56 kDa proteins. The greatest influence of nitric oxide was on the 40 kDa protein: a 17-fold stimulation. Western blotting and immunoprecipitation with specific antibodies identified this protein as the alpha-subunit of G(i-1). Studies with inhibitors showed that the protein was mono-ADP-ribosylated. Treatment of (32P)NAD-labeled G(i-1) with Hg and analysis of the released radioactive material showed that the protein was ADP-ribosylated on a cysteine residue.
与(腺苷酸 - 32P)NAD一起孵育时,有五种睫状体膜蛋白被标记。一氧化氮供体刺激了64、40以及29 - 30 kDa蛋白的标记,并抑制了58和56 kDa蛋白的标记。一氧化氮对40 kDa蛋白的影响最大:刺激了17倍。用特异性抗体进行的蛋白质印迹法和免疫沉淀法鉴定该蛋白为G(i - 1)的α亚基。用抑制剂进行的研究表明该蛋白被单ADP - 核糖基化。用汞处理(32P)NAD标记的G(i - 1)并分析释放的放射性物质表明该蛋白在一个半胱氨酸残基上被ADP - 核糖基化。