Protein partners of dynamin-1 in the retina.
Protein partners of dynamin-1 in the retina.
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DOI：
10.1017/s0952523813000138
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发表时间：
2013-07
影响因子：
1.9
通讯作者：
Hagstrom, Stephanie A.
中科院分区：
文献类型：
--
作者：
Grossman, Gregory H.;Ebke, Lindsey A.;Beight, Craig D.;Jang, Geeng-Fu;Crabb, John W.;Hagstrom, Stephanie A.
发动蛋白参与囊泡的生成，提供机械力以从细胞隔室的膜切除新形成的囊泡。在大脑中，动力蛋白-1、-2和-3已经得到了很好的研究;然而，它们在视网膜中的功能仍然是难以捉摸的。视网膜特异性剪接变体发动蛋白-1与光感受器特异性蛋白质Tubby样蛋白1（Tulp 1）相互作用，当突变时会导致常染色体隐性视网膜色素变性的早发性形式。在这里，我们研究了动力蛋白在视网膜中的作用，使用免疫组织化学定位动力蛋白-1，2和3和免疫沉淀，然后用质谱法探索动力蛋白-1相互作用的蛋白在小鼠视网膜。动力蛋白-2主要局限于光感受器的内节隔室，表明在外节蛋白质运输中的作用。动力蛋白-3存在于光感受器的末端和二级神经元的树突中，但在二级神经元中继来自光感受器的信号的内部丛状层中最明显。动力蛋白-1似乎是视网膜中的主要同种型，并且存在于整个视网膜和感光细胞的多个隔室中。这表明它可能在多种细胞通路中发挥作用。令人惊讶的是，tulp 1 −/−小鼠中发动蛋白-1的表达和定位似乎没有被破坏。免疫沉淀实验表明，发动蛋白-1协会主要与蛋白质参与细胞膜动力学。这一发现通过蛋白质印迹分析得到证实。结果进一步牵连dynamin-1在囊泡蛋白质运输过程中相关的突触和后高尔基体途径，并表明在感光细胞的稳定性可能发挥作用。
Dynamin proteins are involved in vesicle generation, providing mechanical force to excise newly formed vesicles from membranes of cellular compartments. In the brain, dynamin-1, -2 and -3 have been well-studied; however, their function in the retina remains elusive. A retina-specific splice-variant of dynamin-1 interacts with the photoreceptor-specific protein Tubby-like Protein 1 (Tulp1), which when mutated causes an early-onset form of autosomal recessive retinitis pigmentosa. Here we investigated the role of the dynamins in the retina, using immunohistochemistry to localize dynamin-1, -2 and -3 and immunoprecipitation followed by mass spectrometry to explore dynamin-1 interacting proteins in mouse retina. Dynamin-2 is primarily confined to the inner segment compartment of photoreceptors, suggesting a role in outer segment protein transport. Dynamin-3 is present in the terminals of photoreceptors and dendrites of second-order neurons, but is most pronounced in the inner plexiform layer where second-order neurons relay signals from photoreceptors. Dynamin-1 appears to be the dominant isoform in the retina and is present throughout the retina and in multiple compartments of the photoreceptor cell. This suggests that it may function in multiple cellular pathways. Surprisingly, dynamin-1 expression and localization did not appear to be disrupted in tulp1−/− mice. Immunoprecipitation experiments reveal that dynamin-1 associates primarily with proteins involved in cytoskeletal-based membrane dynamics. This finding is confirmed by western blot analysis. Results further implicate dynamin-1 in vesicular protein transport processes relevant to synaptic and post-Golgi pathways and indicate a possible role in photoreceptor stability.