The Fourth Transmembrane Segment Forms the Interface of the Dopamine D2 Receptor Homodimer*

Considerable evidence suggests that G-protein-coupled receptors form homomeric and heteromeric dimersin vivo. Unraveling the structural mechanism for cross-talk between receptors in a dimeric complex must start with the identification of the presently unknown dimer interface. Here, by using cysteine cross-linking, we identify the fourth transmembrane segment (TM4) as a symmetrical dimer interface in the dopamine D2 receptor. Cross-linking is unaffected by ligand binding, and ligand binding and receptor activation are unaffected by cross-linking, suggesting that the receptor is a constitutive dimer. The accessibility of adjacent residues in TM4, however, is affected by ligand binding, implying that the interface has functional significance.

[1]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[2]  J. Falke,et al.  Thermal motions of surface alpha-helices in the D-galactose chemosensory receptor. Detection by disulfide trapping. , 1992, Journal of molecular biology.

[3]  H. Kaback,et al.  A general method for determining helix packing in membrane proteins in situ: helices I and II are close to helix VII in the lactose permease of Escherichia coli. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Michel Bouvier,et al.  A Peptide Derived from a β2-Adrenergic Receptor Transmembrane Domain Inhibits Both Receptor Dimerization and Activation* , 1996, The Journal of Biological Chemistry.

[5]  P. Seeman,et al.  Dopamine D2 receptor dimers and receptor-blocking peptides. , 1996, Biochemical and biophysical research communications.

[6]  C. Romano,et al.  Metabotropic Glutamate Receptor 5 Is a Disulfide-linked Dimer* , 1996, The Journal of Biological Chemistry.

[7]  E. Brown,et al.  Dimerization of the Extracellular Calcium-sensing Receptor (CaR) on the Cell Surface of CaR-transfected HEK293 Cells* , 1998, The Journal of Biological Chemistry.

[8]  Alan Wise,et al.  Heterodimerization is required for the formation of a functional GABAB receptor , 1998, Nature.

[9]  R. Horlick,et al.  Macrophage scavenger receptor confers an adherent phenotype to cells in culture. , 1998, BioTechniques.

[10]  Kenneth A. Jones,et al.  GABAB receptors function as a heteromeric assembly of the subunits GABABR1 and GABABR2 , 1998, Nature.

[11]  R. Shigemoto,et al.  GABAB-receptor subtypes assemble into functional heteromeric complexes , 1998, Nature.

[12]  C Higgs,et al.  Domain swapping in G-protein coupled receptor dimers. , 1998, Protein engineering.

[13]  P. Seeman,et al.  Dopamine D2 receptor dimers in human and rat brain , 1998, FEBS letters.

[14]  Christopher A Reynolds,et al.  Dimerization and Domain Swapping in G-Protein-Coupled Receptors: A Computational Study , 2000, Neuropsychopharmacology.

[15]  J. Javitch,et al.  Amphetamine-induced loss of human dopamine transporter activity: an internalization-dependent and cocaine-sensitive mechanism. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[16]  T. Gudermann,et al.  Structural Implication for Receptor Oligomerization from Functional Reconstitution Studies of Mutant V2 Vasopressin Receptors* , 2000, The Journal of Biological Chemistry.

[17]  H. Akil,et al.  Inhibition of cell surface expression by mutant receptors demonstrates that D2 dopamine receptors exist as oligomers in the cell. , 2000, Molecular pharmacology.

[18]  J. Ballesteros,et al.  The Forgotten Serine , 2000, The Journal of Biological Chemistry.

[19]  Y. Jan,et al.  A Trafficking Checkpoint Controls GABAB Receptor Heterodimerization , 2000, Neuron.

[20]  P. Strange,et al.  Dopamine D2 Receptor Dimer Formation , 2001, The Journal of Biological Chemistry.

[21]  J. Javitch,et al.  Symmetrical dimer of the human dopamine transporter revealed by cross-linking Cys-306 at the extracellular end of the sixth transmembrane segment , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Ballesteros,et al.  Structural mimicry in G protein-coupled receptors: implications of the high-resolution structure of rhodopsin for structure-function analysis of rhodopsin-like receptors. , 2001, Molecular pharmacology.

[23]  Georgios G. Gkoutos,et al.  Lipid-facing correlated mutations and dimerization in G-protein coupled receptors. , 2001, Protein engineering.

[24]  L. Devi,et al.  G-protein-coupled receptor dimerization: modulation of receptor function. , 2001, Pharmacology & therapeutics.

[25]  B. Gowen,et al.  Three-dimensional structure of an invertebrate rhodopsin and basis for ordered alignment in the photoreceptor membrane. , 2001, Journal of molecular biology.

[26]  Susan R. George,et al.  G-Protein-coupled receptor oligomerization and its potential for drug discovery , 2002, Nature Reviews Drug Discovery.

[27]  Jean-François Mercier,et al.  Quantitative Assessment of β1- and β2-Adrenergic Receptor Homo- and Heterodimerization by Bioluminescence Resonance Energy Transfer* , 2002, The Journal of Biological Chemistry.

[28]  L. Prézeau,et al.  The intracellular loops of the GB2 subunit are crucial for G-protein coupling of the heteromeric gamma-aminobutyrate B receptor. , 2002, Molecular pharmacology.

[29]  Michel Bouvier,et al.  Dimerization: an emerging concept for G protein-coupled receptor ontogeny and function. , 2002, Annual review of pharmacology and toxicology.

[30]  Harel Weinstein,et al.  Three-dimensional representations of G protein-coupled receptor structures and mechanisms. , 2002, Methods in enzymology.

[31]  J. Wess,et al.  Use of an in situ disulfide cross-linking strategy to map proximities between amino acid residues in transmembrane domains I and VII of the M3 muscarinic acetylcholine receptor. , 2002, Biochemistry.

[32]  Marta Filizola,et al.  Structural models for dimerization of G-protein coupled receptors: the opioid receptor homodimers. , 2002, Biopolymers.

[33]  Julie Perroy,et al.  A Single Subunit (GB2) Is Required for G-protein Activation by the Heterodimeric GABAB Receptor* , 2002, The Journal of Biological Chemistry.

[34]  K. Palczewski,et al.  Crystal Structure of Rhodopsin: A G‐Protein‐Coupled Receptor , 2002, Chembiochem : a European journal of chemical biology.

[35]  Marta Filizola,et al.  Prediction of heterodimerization interfaces of G-protein coupled receptors with a new subtractive correlated mutation method. , 2002, Protein engineering.