Elucidation of the role of peptide linker in calcium-sensing receptor activation process

被引:15
作者
Ray, Kausik [1 ]
Adipietro, Kaylin A. [1 ]
Chen, Claudia [1 ]
Northup, John K. [1 ]
机构
[1] NIDCD, Lab Cellular Biol, NIH, Bethesda, MD 20892 USA
关键词
D O I
10.1074/jbc.M609610200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Family 3 G-protein-coupled receptors (GPCRs), which includes metabotropic glutamate receptors (mGluRs), sweet and "umami" taste receptors (T1Rs), and the extracellular calcium-sensing receptor (CaR), represent a distinct group among the superfamily of GPCRs characterized by large amino-terminal extracellular ligand-binding domains (ECD) with homology to bacterial periplasmic amino acid-binding proteins that are responsible for signal detection and receptor activation through as yet unresolved mechanism(s) via the seven-transmembrane helical domain (7TMD) common to all GPCRs. To address the mechanism(s) by which ligand-induced conformational changes are conveyed from the ECD to the 7TMD for G-protein activation, we altered the length and composition of a 14-amino acid linker segment common to all family 3 GPCRs except GABA(B) receptor, in the CaR by insertion, deletion, and site-directed mutagenesis of specific highly conserved residues. Small alterations in the length and composition of the linker impaired cell surface expression and abrogated signaling of the chimeric receptors. The exchange of nine amino acids within the linker of CaR with the homologous sequence of mGluR1, however, preserved receptor function. Ala substitution for the four highly conserved residues within this amino acid sequence identified a Len at position 606 of the CaR critical for cell surface expression and signaling. Substitution of Leu(606) for Ala resulted in impaired cell surface expression. However, Ile and Val substitutions displayed strong activating phenotypes. Disruption of the linker by insertion of nine amino acids of a random-coiled structure uncoupled the ECD from regulating the 7TMD. These data are consistent with a model of receptor activation in which the peptide linker, and particularly Leu(606), provides a critical interaction for the CaR signal transmission, a finding likely to be relevant for all family 3 GPCRs containing this conserved motif.
引用
收藏
页码:5310 / 5317
页数:8
相关论文
共 21 条
  • [1] Expression and characterization of inactivating and activating mutations in the human Ca-0(2+)-sensing receptor
    Bai, M
    Quinn, S
    Trivedi, S
    Kifor, O
    Pearce, SHS
    Pollak, MR
    Krapcho, K
    Hebert, SC
    Brown, EM
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (32) : 19537 - 19545
  • [2] Extracellular calcium sensing and extracellular calcium signaling
    Brown, EM
    MacLeod, RJ
    [J]. PHYSIOLOGICAL REVIEWS, 2001, 81 (01) : 239 - 297
  • [3] Allosteric interactions between GB1 and GB2 subunits are required for optimal GABAB receptor function
    Galvez, T
    Duthey, B
    Kniazeff, J
    Blahos, J
    Rovelli, G
    Bettler, B
    Prézeau, L
    Pin, JP
    [J]. EMBO JOURNAL, 2001, 20 (09) : 2152 - 2159
  • [4] Heptahelical domain of metabotropic glutamate receptor 5 behaves like rhodopsin-like receptors
    Goudet, C
    Gaven, F
    Kniazeff, J
    Vol, C
    Liu, JF
    Cohen-Gonsaud, M
    Achers, F
    Prézeau, L
    Pin, JP
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (01) : 378 - 383
  • [5] Hammerland LG, 1999, MOL PHARMACOL, V55, P642
  • [6] Effects of a calcimimetic compound and naturally activating mutations on the human Ca2+ receptor and on Ca2+ receptor/metabotropic glutamate chimeric receptors
    Hauache, OM
    Hu, JX
    Ray, K
    Xie, RY
    Jacobson, KA
    Spiegel, AM
    [J]. ENDOCRINOLOGY, 2000, 141 (11) : 4156 - 4163
  • [7] Human Ca2+ receptor cysteine-rich domain -: Analysis of function of mutant and chimeric receptors
    Hu, JX
    Hauache, O
    Spiegel, AM
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (21) : 16382 - 16389
  • [8] Structural basis of glutamate recognition by a dimeric metabotropic glutamate receptor
    Kunishima, N
    Shimada, Y
    Tsuji, Y
    Sato, T
    Yamamoto, M
    Kumasaka, T
    Nakanishi, S
    Jingami, H
    Morikawa, K
    [J]. NATURE, 2000, 407 (6807) : 971 - 977
  • [9] Ligand-induced signal transduction within heterodimeric GABAB receptor
    Margeta-Mitrovic, M
    Jan, YN
    Jan, LY
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (25) : 14643 - 14648
  • [10] GABAB receptors -: the first 7TM heterodimers
    Marshall, FH
    Jones, KA
    Kaupmann, K
    Bettler, B
    [J]. TRENDS IN PHARMACOLOGICAL SCIENCES, 1999, 20 (10) : 396 - 399