Design of an Ultrafast G Protein Switch Based on a Mouse Melanopsin Variant. (18th June 2019)

• Record Type: Journal Article
• Authors: Tennigkeit, Stefan Alexander; Karapinar, Raziye; Rudack, Till; Dreier, Max‐Aylmer; Althoff, Philipp; Eickelbeck, Dennis; Surdin, Tatjana; Grömmke, Michelle; Mark, Melanie D.; Spoida, Katharina; Lübben, Mathias; Höweler, Udo; Herlitze, Stefan; Gerwert, Klaus
• Is Part Of: Chembiochem. Volume 20:Number 14(2019)
• Page Start: 1766
• Page End: 1771
• Publication Date: 2019-06-18
• Subjects: computational chemistry -- electrophysiology -- integrative modeling -- mutagenesis -- structural biology; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201900110
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: The primary goal of optogenetics is the light‐controlled noninvasive and specific manipulation of various cellular processes. Herein, we present a hybrid strategy for targeted protein engineering combining computational techniques with electrophysiological and UV/visible spectroscopic experiments. We validated our concept for channelrhodopsin‐2 and applied it to modify the less‐well‐studied vertebrate opsin melanopsin. Melanopsin is a promising optogenetic tool that functions as a selective molecular light switch for G protein‐coupled receptor pathways. Thus, we constructed a model of the melanopsin Gq protein complex and predicted an absorption maximum shift of the Y211F variant. This variant displays a narrow blue‐shifted action spectrum and twofold faster deactivation kinetics compared to wild‐type melanopsin on G protein‐coupled inward rectifying K + (GIRK) channels in HEK293 cells. Furthermore, we verified the in vivo activity and optogenetic potential for the variant in mice. Thus, we propose that our developed concept will be generally applicable to designing optogenetic tools. Abstract : A hybrid strategy for targeted protein engineering combines computational chemistry with electrophysiological and UV/Vis spectroscopic experiments. Using this strategy, we designed and characterized a site‐specific variant of the molecular light switch melanopsin that, in combination with other vertebrate opsins, makes a promising optogenetic tool for selective control of two different G protein pathways.