Dimerization deficiency of enigmatic retinitis pigmentosa-linked rhodopsin mutants

• Published in: Nature communications Vol. 7; no. 1; p. 12832
• Main Authors: Ploier, Birgit, Caro, Lydia N., Morizumi, Takefumi, Pandey, Kalpana, Pearring, Jillian N., Goren, Michael A., Finnemann, Silvia C., Graumann, Johannes, Arshavsky, Vadim Y., Dittman, Jeremy S., Ernst, Oliver P., Menon, Anant K.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 03.10.2016; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 14/35; 631/114/2411; 631/45/612/194; 631/57/2283; 82/1; 82/16; 82/29; 82/51; 82/80; 82/83; Animals; Biochemistry; Cattle; Chlorocebus aethiops; COS Cells; Disease; GTP-Binding Proteins - chemistry; HEK293 Cells; Humanities and Social Sciences; Humans; Liposomes - metabolism; Membranes; Mice, Knockout; multidisciplinary; Mutation; Phospholipid Transfer Proteins - metabolism; Photoreceptors; Pigments; Point Mutation; Protein Multimerization; Proteins; Retina - chemistry; Retina - metabolism; Retinitis Pigmentosa - genetics; Rhodopsin - chemistry; Rhodopsin - genetics; Science; Science (multidisciplinary); Transducin - genetics; North Carolina; New York; Canada; Toronto Ontario Canada; United States > US; Qatar
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms12832

Retinitis pigmentosa (RP) is a blinding disease often associated with mutations in rhodopsin, a light-sensing G protein-coupled receptor and phospholipid scramblase. Most RP-associated mutations affect rhodopsin’s activity or transport to disc membranes. Intriguingly, some mutations produce apparently normal rhodopsins that nevertheless cause disease. Here we show that three such enigmatic mutations—F45L, V209M and F220C—yield fully functional visual pigments that bind the 11- cis retinal chromophore, activate the G protein transducin, traffic to the light-sensitive photoreceptor compartment and scramble phospholipids. However, tests of scramblase activity show that unlike wild-type rhodopsin that functionally reconstitutes into liposomes as dimers or multimers, F45L, V209M and F220C rhodopsins behave as monomers. This result was confirmed in pull-down experiments. Our data suggest that the photoreceptor pathology associated with expression of these enigmatic RP-associated pigments arises from their unexpected inability to dimerize via transmembrane helices 1 and 5. Retinitis pigmentosa is often caused by mutations that affect the activity or transport of rhodopsin, but some mutations cause disease even though an apparently functional protein is produced. Here the authors show that three such enigmatic mutants retain scramblase activity but are unable to dimerize.