Kinetics of the early events of GPCR signalling

被引:12
作者
Adamson, Roslin J. [1 ]
Watts, Anthony [1 ]
机构
[1] Univ Oxford, Dept Biochem, Biomembrane Struct Unit, Oxford OX1 3QU, England
基金
英国惠康基金;
关键词
G protein-coupled receptor; Electron microscopy; Surface plasmon resonance; Nanodisc; G protein; PROTEIN-COUPLED RECEPTORS; DELTA-OPIOID RECEPTOR; MEMBRANE-PROTEINS; PLASMON RESONANCE; LIGAND-BINDING; NEUROTENSIN; NANODISCS; RECONSTITUTION; PURIFICATION; RHODOPSIN;
D O I
10.1016/j.febslet.2014.10.043
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
Neurotensin receptor type 1 (NTS1) is a G protein-coupled receptor (GPCR) that affects cellular responses by initiating a cascade of interactions through G proteins. The kinetic details for these interactions are not well-known. Here, NTS1-nanodisc-G alpha(s) and G alpha(i1) interactions were studied. The binding affinities of G alpha(i1) and G alpha(s) to NTS1 were directly measured by surface plasmon resonance (SPR) and determined to be 15 +/- 6 nM and 31 +/- 18 nM, respectively. This SPR configuration permits the kinetics of early events in signalling pathways to be explored and can be used to initiate descriptions of the GPCR interactome. (C) 2014 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
引用
收藏
页码:4701 / 4707
页数:7
相关论文
共 61 条
[11]   Directed self-assembly of monodisperse phospholipid bilayer nanodiscs with controlled size [J].
Denisov, IG ;
Grinkova, YV ;
Lazarides, AA ;
Sligar, SG .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (11) :3477-3487
[12]   Examining the contributions of lipid shape and headgroup charge on bilayer behavior [J].
Dickey, Allison ;
Faller, Roland .
BIOPHYSICAL JOURNAL, 2008, 95 (06) :2636-2646
[13]   Neurotensin and pain modulation [J].
Dobner, Paul R. .
PEPTIDES, 2006, 27 (10) :2405-2414
[14]   The G protein subunit gene families [J].
Downes, GB ;
Gautam, N .
GENOMICS, 1999, 62 (03) :544-552
[15]   Endogenous CNS Expression of Neurotensin and Neurotensin Receptors Is Altered during the Postpartum Period in Outbred Mice [J].
Driessen, Terri M. ;
Zhao, Changjiu ;
Whittlinger, Anna ;
Williams, Horecia ;
Gammie, Stephen C. .
PLOS ONE, 2014, 9 (01)
[16]   Nanodiscs allow the use of integral membrane proteins as analytes in surface plasmon resonance studies [J].
Glueck, Julian M. ;
Koenig, Bernd W. ;
Willbold, Dieter .
ANALYTICAL BIOCHEMISTRY, 2011, 408 (01) :46-52
[17]  
Greentree Wendy K., 2003, V237, P3
[18]   Functional coupling with Gαq and Gαil protein subunits promotes high-affinity agonist binding to the neurotensin receptor NTS-1 expressed in Escherichia coli [J].
Grisshammer, R ;
Hermans, E .
FEBS LETTERS, 2001, 493 (2-3) :101-105
[19]  
Grisshammer Reinhard, 2005, Journal of Structural and Functional Genomics, V6, P159, DOI 10.1007/s10969-005-1917-6
[20]   Neurotensin receptor type 1:: Escherichia coli expression, purification, characterization and biophysical study [J].
Harding, P. J. ;
Attrill, H. ;
Ross, S. ;
Koeppe, J. R. ;
Kapanidis, A. N. ;
Watts, A. .
BIOCHEMICAL SOCIETY TRANSACTIONS, 2007, 35 :760-763