Functional expression in vitro of bovine visual rhodopsin.
Expression of functionally active bovine visual rhodopsin was achieved by sequential transcription and translation in vitro of rhodopsin gene cDNA with co-translational insertion of the protein into phosphatidylcholine liposomes. The recombinant rhodopsin has functional, spectral and immunochemical properties similar to those of natural rhodopsin from bovine retina. Two mutant rhodopsins, Cys316----Ser and Asp330----Asn, Asp331----Asn, were produced by oligonucleotide-directed mutagenesis. The first mutation does not affect rhodopsin's ability to stimulate transducin GTPase and visual cGMP phosphodiesterase activities, while the second double mutation leads to a sharp decrease in rhodopsin activity.
Zozulya
S A
SA
Branch of M.M.Shemyakin Institute of Bioorganic Chemistry, Academy of Sciences, Pushchino, Moscow Region, USSR.
Gurevich
V V
VV
Zvyaga
T A
TA
Shirokova
E P
EP
Dumler
I L
IL
Garnovskaya
M N
MN
Natochin MYu
Shmukler
B E
BE
Badalov
P R
PR
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics; D015870:Gene Expression; D005796:Genes; D008969:Molecular Sequence Data; D009154:Mutation; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012168:Retinal Pigments / Q000235:genetics*; D012243:Rhodopsin / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D017299:Rod Opsins; D014158:Transcription, Genetic
D005136:Eye Proteins; D011994:Recombinant Proteins; D012168:Retinal Pigments; D017299:Rod Opsins; D004247:DNA; D012243:Rhodopsin