From the Department of Biological Sciences, Graduate School of Science, and.
J Biol Chem. 2014 Jan 31;289(5):2651-7. doi: 10.1074/jbc.M113.495325. Epub 2013 Dec 16.
Cones are less light-sensitive than rods. We showed previously in carp that more light (>100-fold) is required in cones than in rods to activate 50% of cGMP phosphodiesterase (PDE). The lower effectiveness of PDE activation in carp cones is due partly to the fact that the activation rate of transducin (Tr) by light-activated visual pigment (R*) is 5-fold lower in carp cones than in rods. In this study, we tried to explain the remaining difference. First, we examined the efficiency of activation of PDE by activated Tr (Tr*). By activating PDE with known concentrations of the active (guanosine 5'-Ο-(γ-thio)triphosphate (GTPγS)-bound) form of Tr*, we found that Tr* activated PDE at a similar efficiency in rods and cones. Next, we examined the contribution of R* and Tr* lifetimes. In a comparison of PDE activation in the presence (with GTP) and absence (with GTPγS) of Tr* inactivation, PDE activation required more light (and was therefore less effective) when Tr* was inactivated in both rod and cone membranes. This is probably because inactivation of Tr* shortened its lifetime, thereby reducing the number of activated PDE molecules. The effect of Tr* inactivation was larger in cones, probably because the lifetime of Tr* is shorter in cones than in rods. The shorter lifetimes of Tr* and R* in cones seem to explain the remaining difference in the effectiveness of PDE activation between rods and cones.
视锥细胞比视杆细胞对光的敏感度低。我们之前在鲤鱼中表明,激活 50%的环鸟苷酸磷酸二酯酶(PDE)所需的光量(>100 倍)在视锥细胞中比在视杆细胞中更多。鲤鱼视锥细胞中 PDE 激活效率较低的部分原因是,光激活视觉色素(R*)激活转导蛋白(Tr)的激活速率在鲤鱼视锥细胞中比在视杆细胞中低 5 倍。在这项研究中,我们试图解释其余的差异。首先,我们检查了激活的 Tr(Tr*)对 PDE 的激活效率。通过用已知浓度的活性(鸟苷 5'-O-(γ-硫)三磷酸(GTPγS)结合)形式的 Tr激活 PDE,我们发现 Tr在视杆细胞和视锥细胞中以相似的效率激活 PDE。接下来,我们检查了 R和 Tr寿命的贡献。在比较 PDE 在有(含 GTP)和无(含 GTPγS)Tr失活的情况下的激活,当 Tr在视杆和视锥细胞膜中都失活时,PDE 的激活需要更多的光(因此效率更低)。这可能是因为 Tr的失活缩短了它的寿命,从而减少了激活的 PDE 分子的数量。Tr失活的影响在视锥细胞中更大,可能是因为 Tr在视锥细胞中的寿命比在视杆细胞中更短。Tr和 R*在视锥细胞中的较短寿命似乎解释了视杆细胞和视锥细胞中 PDE 激活效率之间剩余差异的原因。