Home LiteratureArticle Details
PMID: 3141920 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Expression of a bovine rhodopsin gene in Xenopus oocytes: demonstration of light-dependent ionic currents.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 21 ·1988-11-00 ·Pages 7917-21

Khorana HG, Knox BE, Nasi E, Swanson R, Thompson DA

Abstract

Xenopus oocytes express a gene encoding bovine rhodopsin as well as its SP6 RNA polymerase-derived transcripts and total retinal mRNA. The opsin produced is in unglycosylated (30 kDa) and two glycosylated (35 kDa and 41 kDa) forms. Incubation of the cells expressing the above proteins with 11-cis-retinal generates rhodopsin, which was purified by immunoaffinity chromatography. The purified protein shows the expected UV/visible absorption spectrum and characteristic light-dependent activation of the rod outer segment GTP binding protein. Oocytes expressing rhodopsin exhibit light-dependent ionic currents that are detected by voltage-clamp techniques.

MeSH Terms
Animals Cattle GTP-Binding Proteins/metabolism Glycosylation Light Mutation Oocytes/metabolism RNA, Messenger/analysis Retina/analysis Retinal Pigments/genetics Rhodopsin/genetics Xenopus
Chemicals
RNA, Messenger Retinal Pigments Rhodopsin GTP-Binding Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Khorana H G
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Knox B E
Nasi E
Swanson R
Thompson D A
References (33)
33 references, click to expand
  1. A method for preparing stable digitonin solutions for visual pigment extraction.
    Vision Res. 1977 Feb;17(2):301-2 PMID: 867852
  2. Synthesis and bleaching of rhodopsin.
    Nature. 1956 Jan 28;177(4500):174-6 PMID: 13297008
  3. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
    Biochemistry. 1979 Nov 27;18(24):5294-9 PMID: 518835
  4. Characterization of bovine rod outer segment G-protein.
    J Biol Chem. 1982 Jun 10;257(11):6452-60 PMID: 7076677
  5. A transient calcium-dependent chloride current in the immature Xenopus oocyte.
    J Physiol. 1983 Sep;342:309-25 PMID: 6313909
  6. N-methyl-1-deoxynojirimycin, a novel inhibitor of glycoprotein processing, and its effect on fowl plague virus maturation.
    Virology. 1983 Oct 15;130(1):238-42 PMID: 6636538
  7. The molecular basis of visual excitation.
    Nature. 1968 Aug 24;219(5156):800-7 PMID: 4876934
  8. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  9. Purification and characterization of adenosine triphosphate: ribonucleic acid adenyltransferase from Escherichia coli.
    Eur J Biochem. 1973 Aug 1;37(1):31-40 PMID: 4580885
  10. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus.
    J Biol Chem. 1974 Feb 10;249(3):811-7 PMID: 4204552
  11. Novel mannosidase inhibitor blocking conversion of high mannose to complex oligosaccharides.
    Nature. 1984 Feb 23-29;307(5953):755-8 PMID: 6230538
  12. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  13. Recognition of cap structure in splicing in vitro of mRNA precursors.
    Cell. 1984 Oct;38(3):731-6 PMID: 6567484
  14. Specificity of the functional interactions of the beta-adrenergic receptor and rhodopsin with guanine nucleotide regulatory proteins reconstituted in phospholipid vesicles.
    J Biol Chem. 1985 Feb 10;260(3):1493-500 PMID: 2981858
  15. The effect of capping and polyadenylation on the stability, movement and translation of synthetic messenger RNAs in Xenopus oocytes.
    Nucleic Acids Res. 1985 Oct 25;13(20):7375-94 PMID: 3932972
  16. Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F.
    Biochemistry. 1985 Aug 13;24(17):4665-71 PMID: 4063349
  17. Total synthesis of a gene for bovine rhodopsin.
    Proc Natl Acad Sci U S A. 1986 Feb;83(3):599-603 PMID: 3456156
  18. Cyclic GMP cascade of vision.
    Annu Rev Neurosci. 1986;9:87-119 PMID: 2423011
  19. Cloning, sequencing and expression of complementary DNA encoding the muscarinic acetylcholine receptor.
    Nature. 1986 Oct 2-8;323(6087):411-6 PMID: 3762692
  20. Involvement of a GTP-binding protein in mediation of serotonin and acetylcholine responses in Xenopus oocytes injected with rat brain messenger RNA.
    Brain Res. 1986 Dec;387(3):201-9 PMID: 2435359
  21. Multiple opsin mRNA species in bovine retina.
    Brain Res. 1986 Dec;387(3):251-60 PMID: 2950966
  22. cDNA cloning of a serotonin 5-HT1C receptor by electrophysiological assays of mRNA-injected Xenopus oocytes.
    Proc Natl Acad Sci U S A. 1987 Jun;84(12):4332-6 PMID: 3473504
  23. Rat brain serotonin receptors in Xenopus oocytes are coupled by intracellular calcium to endogenous channels.
    Proc Natl Acad Sci U S A. 1987 Jul;84(14):5063-7 PMID: 2440042
  24. A family of receptors coupled to guanine nucleotide regulatory proteins.
    Biochemistry. 1987 May 19;26(10):2657-64 PMID: 3038163
  25. Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains.
    Annu Rev Biochem. 1987;56:497-534 PMID: 3304143
  26. G proteins: transducers of receptor-generated signals.
    Annu Rev Biochem. 1987;56:615-49 PMID: 3113327
  27. Inositol phosphate formation and chloride current responses induced by acetylcholine and serotonin through GTP-binding proteins in Xenopus oocyte after injection of rat brain messenger RNA.
    Brain Res. 1987 Jul;388(2):113-23 PMID: 2887235
  28. cDNA cloning of bovine substance-K receptor through oocyte expression system.
    Nature. 1987 Oct 29-Nov 4;329(6142):836-8 PMID: 2823146
  29. Expression of a synthetic bovine rhodopsin gene in monkey kidney cells.
    Proc Natl Acad Sci U S A. 1987 Dec;84(24):8874-8 PMID: 2962193
  30. A single amino acid substitution in rhodopsin (lysine 248----leucine) prevents activation of transducin.
    J Biol Chem. 1988 Feb 15;263(5):2119-22 PMID: 3123487
  31. The use of Xenopus oocytes for the study of ion channels.
    CRC Crit Rev Biochem. 1987;22(4):317-87 PMID: 2449311
  32. Fertilization events induced by neurotransmitters after injection of mRNA in Xenopus eggs.
    Science. 1988 Jul 22;241(4864):464-7 PMID: 3134693
  33. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.21.7917
Published
1988-11-00
Pages
7917-21
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC282324
Subset
IM
Grants
NEI NIH HHS · 1F32EY05916 · United States
NEI NIH HHS · 1F32EY06024 · United States
NIGMS NIH HHS · GM28289-08 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.21.7917 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2