Chimeric human opsins as optogenetic light sensitisers.
Chimeric human opsins as optogenetic light sensitisers.
复制标题
DOI：
10.1242/jeb.240580
复制
发表时间：
2021-07-15
期刊：
影响因子：
--
通讯作者：
Hankins MW
中科院分区：
文献类型：
--
作者：
Hickey DG;Davies WIL;Hughes S;Rodgers J;Thavanesan N;MacLaren RE;Hankins MW
人类视蛋白为基础的色素作为光敏剂有很大的潜力，但他们的光转导级联特异性第二信使蛋白的要求可能会限制其在非天然细胞类型的功能。在这项研究中，产生了八种嵌合的人视蛋白，其由视紫红质（RHO）或长波长敏感（LWS）视蛋白的骨架和来自Gq/11偶联的人黑视蛋白的细胞内结构域组成。与Gi和Gq/11途径偶联的视紫红质/黑视蛋白嵌合视蛋白。用相应的黑视素结构域对细胞内表面的更大取代通常显示出更大的Gq/11活性，而Gi活化降低。与黑视素不同，视紫红质和视紫红质/黑视素嵌合体依赖于外源性发色团发挥功能。相比之下，野生型LWS视蛋白和LWS视蛋白/黑视蛋白嵌合体在对光的反应中仅显示出弱的GI激活，而未检测到Gq/11途径激活。免疫细胞化学（ICC）表明，嵌合视蛋白与更多的黑视蛋白的细胞内结构域不太可能被贩运到质膜。这项研究证明了Gα偶联效率对细胞反应速度的重要性，并创造了具有独特特性组合的人类视蛋白，以扩大用于基础研究和转化疗法的定制光遗传学生物工具的范围。总结：将人类视觉视蛋白和黑视蛋白的不同结构域组合产生具有潜在光遗传学应用的功能独特的嵌合视蛋白。
Human opsin-based photopigments have great potential as light-sensitisers, but their requirement for phototransduction cascade-specific second messenger proteins may restrict their functionality in non-native cell types. In this study, eight chimeric human opsins were generated consisting of a backbone of either a rhodopsin (RHO) or long-wavelength-sensitive (LWS) opsin and intracellular domains from Gq/11-coupled human melanopsin. Rhodopsin/melanopsin chimeric opsins coupled to both Gi and Gq/11 pathways. Greater substitution of the intracellular surface with corresponding melanopsin domains generally showed greater Gq/11 activity with a decrease in Gi activation. Unlike melanopsin, rhodopsin and rhodopsin/melanopsin chimeras were dependent upon exogenous chromophore to function. By contrast, wild-type LWS opsin and LWS opsin/melanopsin chimeras showed only weak Gi activation in response to light, whilst Gq/11 pathway activation was not detected. Immunocytochemistry (ICC) demonstrated that chimeric opsins with more intracellular domains of melanopsin were less likely to be trafficked to the plasma membrane. This study demonstrates the importance of Gα coupling efficiency to the speed of cellular responses and created human opsins with a unique combination of properties to expand the range of customised optogenetic biotools for basic research and translational therapies. Summary: Combining different domains of human visual opsins and melanopsin creates functionally unique chimeric opsins with potential optogenetic applications.
登录
查看更多内容
影响因子：
3.7
作者：
Bailes HJ;Zhuang LY;Lucas RJ
通讯作者：
Lucas RJ
影响因子：
56.9
作者：
KOBILKA, BK;KOBILKA, TS;LEFKOWITZ, RJ
通讯作者：
LEFKOWITZ, RJ
影响因子：
2.9
作者：
Kim, JM;Hwa, J;Khorana, HG
通讯作者：
Khorana, HG
影响因子：
16.2
作者：
Emanuel, Alan Joseph;Do, Michael Tri Hoang
通讯作者：
Do, Michael Tri Hoang
影响因子：
8
作者：
Hughes, Steven;Jagannath, Aarti;Hankins, Mark W.
通讯作者：
Hankins, Mark W.