Dimerization deficiency of enigmatic retinitis pigmentosa-linked rhodopsin mutants.
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.
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, New York 10065, USA.
Birgit
B
Ploier
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
Lydia N
LN
Caro
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
Takefumi
T
Morizumi
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, New York 10065, USA.
Kalpana
K
Pandey
Department of Ophthalmology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Jillian N
JN
Pearring
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, New York 10065, USA.
Michael A
MA
Goren
Department of Biological Sciences, Center for Cancer, Genetic Diseases and Gene Regulation, Fordham University, Bronx, New York 10458, USA.
Silvia C
SC
Finnemann
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, New York 10065, USA.|Weill Cornell Medicine-Qatar, Qatar Foundation, Education City P.O.Box 24144, Doha, State of Qatar.
Johannes
0000-0002-3015-5850
J
Graumann
Department of Ophthalmology, Duke University Medical Center, Durham, North Carolina 27710, USA.|Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Vadim Y
VY
Arshavsky
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, New York 10065, USA.
Jeremy S
JS
Dittman
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8.|Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
Oliver P
OP
Ernst
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, New York 10065, USA.
Anant K
0000-0001-6924-2698
AK
Menon
D000818:Animals; D019556:COS Cells; D002417:Cattle; D002522:Chlorocebus aethiops; D019204:GTP-Binding Proteins / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D018345:Mice, Knockout; D009154:Mutation; D049294:Phospholipid Transfer Proteins / Q000378:metabolism; D017354:Point Mutation; D055503:Protein Multimerization; D012160:Retina / Q000737:chemistry / Q000378:metabolism*; D012174:Retinitis Pigmentosa / Q000235:genetics*; D012243:Rhodopsin / Q000737:chemistry* / Q000235:genetics*; D015293:Transducin / Q000235:genetics
D008081:Liposomes; D049294:Phospholipid Transfer Proteins; D012243:Rhodopsin; D019204:GTP-Binding Proteins; D015293:Transducin
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