Caretta A, Stein P J
Biochemistry. 1986 May 6;25(9):2335-41. doi: 10.1021/bi00357a006.
Under conditions in which large guanosine cyclic 3',5'-phosphate (cGMP)- and phosphodiesterase (PDE)-dependent changes in near-infrared transmission and vesicle aggregation and disaggregation occur, we have observed a striking change in the binding of PDE to rod disk membranes. The change in PDE binding is nucleotide and light dependent as are the light-scattering changes. The cGMP- and PDE-dependent light-scattering signal can be produced by a 500-nm light flash which bleaches 1/(1 X 10(7] rhodopsin molecules. Mg ions are an essential cofactor for the nucleotide-dependent PDE binding and light-scattering changes. 3-Isobutyl-1-methylxanthine and other competitive inhibitors of PDE hydrolytic activity support increased PDE binding to the disk membrane, vesicle aggregation, and the light-scattering signal. However, treatments which block GTP-dependent activation of PDE hydrolytic activity (colchicine, GDP, or ethylenediaminetetraacetic acid) also block these phenomena. Thus, GTP-dependent activation of PDE rather than its hydrolytic activity appears to be correlated with the light-scattering signal.
在近红外透射以及囊泡聚集和解聚过程中出现大的鸟苷环化3',5'-磷酸(cGMP)和磷酸二酯酶(PDE)依赖性变化的条件下,我们观察到PDE与视杆盘膜结合发生了显著变化。PDE结合的变化与核苷酸和光有关,光散射变化也是如此。cGMP和PDE依赖性光散射信号可由500nm的光脉冲产生,该光脉冲可使1/(1×10⁷)视紫红质分子漂白。镁离子是核苷酸依赖性PDE结合和光散射变化的必需辅助因子。3-异丁基-1-甲基黄嘌呤和其他PDE水解活性的竞争性抑制剂可支持PDE与盘膜结合增加、囊泡聚集以及光散射信号增强。然而,阻断PDE水解活性的GTP依赖性激活的处理(秋水仙碱、GDP或乙二胺四乙酸)也会阻断这些现象。因此,PDE的GTP依赖性激活而非其水解活性似乎与光散射信号相关。