Home LiteratureArticle Details
PMID: 21925606 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Rhodopsin-transducin heteropentamer: three-dimensional structure and biochemical characterization.

Journal of structural biology ·Vol. 176 ·No. 3 ·2011-12-00 ·Pages 387-94

Jastrzebska B, Ringler P, Lodowski DT, Moiseenkova-Bell V, Golczak M, Müller SA, Palczewski K, Engel A

Abstract

The process of vision is initiated when the G protein-coupled receptor, rhodopsin (Rho), absorbs a photon and transitions to its activated Rho(∗) form. Rho(∗) binds the heterotrimeric G protein, transducin (G(t)) inducing GDP to GTP exchange and G(t) dissociation. Using nucleotide depletion and affinity chromatography, we trapped and purified the resulting nucleotide-free Rho(∗)·G(t) complex. Quantitative SDS-PAGE suggested a 2:1 molar ratio of Rho(∗) to G(t) in the complex and its mass determined by scanning transmission electron microscopy was 221±12kDa. A 21.6Å structure was calculated from projections of negatively stained Rho(∗)·G(t) complexes. The molecular envelope thus determined accommodated two Rho molecules together with one G(t) heterotrimer, corroborating the heteropentameric structure of the Rho(∗)·G(t) complex.

MeSH Terms
Electrophoresis, Polyacrylamide Gel Guanosine Diphosphate/chemistry Guanosine Triphosphate/chemistry Light Microscopy, Electron, Transmission Models, Molecular Multiprotein Complexes/chemistry,ultrastructure Protein Binding Protein Multimerization Rhodopsin/chemistry,ultrastructure Transducin/chemistry,ultrastructure
Chemicals
Multiprotein Complexes Guanosine Diphosphate Guanosine Triphosphate Rhodopsin Transducin
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Jastrzebska Beata
Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4965, USA. bxj27@case.edu
Ringler Philippe
Lodowski David T
Moiseenkova-Bell Vera
Golczak Marcin
Müller Shirley A
Palczewski Krzysztof
Engel Andreas
References (38)
38 references, click to expand
  1. EMAN2: an extensible image processing suite for electron microscopy.
    J Struct Biol. 2007 Jan;157(1):38-46 PMID: 16859925
  2. Organization of the G protein-coupled receptors rhodopsin and opsin in native membranes.
    J Biol Chem. 2003 Jun 13;278(24):21655-21662 PMID: 12663652
  3. Dimerization: an emerging concept for G protein-coupled receptor ontogeny and function.
    Annu Rev Pharmacol Toxicol. 2002;42:409-35 PMID: 11807178
  4. Lateral diffusion of rhodopsin in photoreceptor membrane: a reappraisal.
    Mol Vis. 2009 Aug 28;15:1717-29 PMID: 19727341
  5. Crystal structure of the β2 adrenergic receptor-Gs protein complex.
    Nature. 2011 Jul 19;477(7366):549-55 PMID: 21772288
  6. Crystal structure of a photoactivated deprotonated intermediate of rhodopsin.
    Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16123-8 PMID: 17060607
  7. Transducin interactions with rhodopsin. Evidence for positive cooperative behavior.
    J Biol Chem. 1987 Sep 15;262(26):12444-7 PMID: 3114258
  8. Allosteric communication between protomers of dopamine class A GPCR dimers modulates activation.
    Nat Chem Biol. 2009 Sep;5(9):688-95 PMID: 19648932
  9. The 2.0 A crystal structure of a heterotrimeric G protein.
    Nature. 1996 Jan 25;379(6563):311-9 PMID: 8552184
  10. Role of membrane integrity on G protein-coupled receptors: Rhodopsin stability and function.
    Prog Lipid Res. 2011 Jul;50(3):267-77 PMID: 21435354
  11. Rhodopsin: insights from recent structural studies.
    Annu Rev Biophys Biomol Struct. 2002;31:443-84 PMID: 11988478
  12. G protein-coupled receptor rhodopsin.
    Annu Rev Biochem. 2006;75:743-67 PMID: 16756510
  13. Phospholipids are needed for the proper formation, stability, and function of the photoactivated rhodopsin-transducin complex.
    Biochemistry. 2009 Jun 16;48(23):5159-70 PMID: 19413332
  14. Rhodopsin-interacting surface of the transducin gamma subunit.
    Biochemistry. 2006 Aug 8;45(31):9386-92 PMID: 16878973
  15. The supramolecular structure of the GPCR rhodopsin in solution and native disc membranes.
    Mol Membr Biol. 2004 Nov-Dec;21(6):435-46 PMID: 15764373
  16. Interaction of a G protein with an activated receptor opens the interdomain interface in the alpha subunit.
    Proc Natl Acad Sci U S A. 2011 Jun 7;108(23):9420-4 PMID: 21606326
  17. EMAN: semiautomated software for high-resolution single-particle reconstructions.
    J Struct Biol. 1999 Dec 1;128(1):82-97 PMID: 10600563
  18. UCSF Chimera--a visualization system for exploratory research and analysis.
    J Comput Chem. 2004 Oct;25(13):1605-12 PMID: 15264254
  19. Responses of retinal rods to single photons.
    J Physiol. 1979 Mar;288:613-34 PMID: 112243
  20. Isolation and functional characterization of a stable complex between photoactivated rhodopsin and the G protein, transducin.
    FASEB J. 2009 Feb;23(2):371-81 PMID: 18827025
  21. Efficient coupling of transducin to monomeric rhodopsin in a phospholipid bilayer.
    J Biol Chem. 2008 Feb 15;283(7):4387-94 PMID: 18033822
  22. Crystal structure of rhodopsin: A G protein-coupled receptor.
    Science. 2000 Aug 4;289(5480):739-45 PMID: 10926528
  23. A G protein-coupled receptor at work: the rhodopsin model.
    Trends Biochem Sci. 2009 Nov;34(11):540-52 PMID: 19836958
  24. Interactions between photoexcited rhodopsin and GTP-binding protein: kinetic and stoichiometric analyses from light-scattering changes.
    Proc Natl Acad Sci U S A. 1981 Nov;78(11):6873-7 PMID: 6273893
  25. Crystal structure of metarhodopsin II.
    Nature. 2011 Mar 31;471(7340):651-5 PMID: 21389988
  26. The retinal conformation and its environment in rhodopsin in light of a new 2.2 A crystal structure.
    J Mol Biol. 2004 Sep 10;342(2):571-83 PMID: 15327956
  27. G-protein alpha and beta-gamma subunits interact with conformationally distinct signaling states of rhodopsin.
    Vision Res. 2006 Dec;46(27):4442-8 PMID: 16989885
  28. Single particle reconstructions of the transferrin-transferrin receptor complex obtained with different specimen preparation techniques.
    J Mol Biol. 2006 Feb 3;355(5):1048-65 PMID: 16343539
  29. Mesoscopic Monte Carlo simulations of stochastic encounters between photoactivated rhodopsin and transducin in disc membranes.
    J Phys Chem B. 2008 Apr 10;112(14):4419-26 PMID: 18345658
  30. Transducin activation by nanoscale lipid bilayers containing one and two rhodopsins.
    J Biol Chem. 2007 May 18;282(20):14875-81 PMID: 17395586
  31. Atomic-force microscopy: Rhodopsin dimers in native disc membranes.
    Nature. 2003 Jan 9;421(6919):127-8 PMID: 12520290
  32. A dynamic scaffolding mechanism for rhodopsin and transducin interaction in vertebrate vision.
    Biochem J. 2011 Dec 1;440(2):263-71 PMID: 21843151
  33. Rescue of defective G protein-coupled receptor function in vivo by intermolecular cooperation.
    Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):2319-24 PMID: 20080658
  34. Homodimerization of the beta2-adrenergic receptor as a prerequisite for cell surface targeting.
    J Biol Chem. 2004 Aug 6;279(32):33390-7 PMID: 15155738
  35. Semper: Distortion Compensation, Selective Averaging, 3-D Reconstruction, and Transfer Function Correction in a Highly Programmable System
    J Struct Biol. 1996 Oct;116(1):230-6 PMID: 8812977
  36. The structural basis of agonist-induced activation in constitutively active rhodopsin.
    Nature. 2011 Mar 31;471(7340):656-60 PMID: 21389983
  37. Crystal structure of opsin in its G-protein-interacting conformation.
    Nature. 2008 Sep 25;455(7212):497-502 PMID: 18818650
  38. MASDET-A fast and user-friendly multiplatform software for mass determination by dark-field electron microscopy.
    J Struct Biol. 2009 Feb;165(2):78-87 PMID: 19041401
Article Info
Journal
Journal of structural biology
Abbr.
J Struct Biol
ISSN
1095-8657
DOI
10.1016/j.jsb.2011.08.016
Email
bxj27@case.edu
Published
2011-12-00
Epub
2011-00-06
Pages
387-94
Language
English
Region
United States
NLM ID
9011206
PMCID
PMC3210324
Subset
IM
Grants
NEI NIH HHS · P30 EY011373 · United States
NIGMS NIH HHS · GM079191 · United States
NEI NIH HHS · R01 EY008061 · United States
NEI NIH HHS · R00 EY019718 · United States
NIGMS NIH HHS · R01 GM079191 · United States
NEI NIH HHS · EY019718 · United States
NEI NIH HHS · R01 EY019478-01 · United States
NEI NIH HHS · P30 EY011373-08 · United States
NEI NIH HHS · R01 EY019478 · United States
NEI NIH HHS · EY008061 · United States
NEI NIH HHS · K99 EY019718 · United States
NEI NIH HHS · R01 EY008061-16 · United States
NEI NIH HHS · P30EY11373 · United States
NIGMS NIH HHS · U54 GM094598 · United States
NIGMS NIH HHS · R01 GM079191-01 · United States
External Links
PubMed source DOI: 10.1016/j.jsb.2011.08.016 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: product@genelibs.com

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