Characterization of radioligand binding to a transmembrane receptor reconstituted into Lipobeads

被引:8
作者
Park, PSH
Ng, CC
Buck, S
Wells, JW
Cheng, YL
Pennefather, PS [1 ]
机构
[1] Univ Toronto, Leslie Dan Fac Pharm, Dept Pharmaceut Sci, Toronto, ON M5S 2S2, Canada
[2] Univ Toronto, Dept Chem Engn & Appl Chem, Toronto, ON M5S 3E5, Canada
来源
FEBS LETTERS | 2004年 / 567卷 / 2-3期
关键词
assay system; biomaterial; biosensors and nanotechnology; muscarinic receptor; lipobead; transmembrane receptor;
D O I
10.1016/j.febslet.2004.03.124
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
Lipobeads are hydrogel beads surrounded by a lipid bilayer membrane and have been developed to act as a cell analogue. The FLAG-tagged M-2 muscarinic receptor was incorporated onto the surface of the Lipobead by incubating pre-Lipobeads with proteoliposomes containing the receptor. Receptors reconstituted onto the surface of the Lipobeads were functional in that they bound the antagonists quinuclidinylbenzilate and scopolamine with characteristic muscarinic affinities. This demonstrates the feasibility of using Lipobeads to study the binding properties of the M-2 muscarinic receptor and offers a promising approach to the study of transmembrane protein biology in general. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
引用
收藏
页码:344 / 348
页数:5
相关论文
共 23 条
[1]   Cardiac muscarinic receptors. Cooperativity as the basis for multiple states of affinity [J].
Chidiac, P ;
Green, MA ;
Pawagi, AB ;
Wells, JW .
BIOCHEMISTRY, 1997, 36 (24) :7361-7379
[2]   A biosensor that uses ion-channel switches [J].
Cornell, BA ;
BraachMaksvytis, VLB ;
King, LG ;
Osman, PDJ ;
Raguse, B ;
Wieczorek, L ;
Pace, RJ .
NATURE, 1997, 387 (6633) :580-583
[3]  
DE MI, 2000, PHARM RES, V17, P817
[4]   G protein-coupled receptor microarrays for drug discovery [J].
Fang, Y ;
Lahiri, J ;
Picard, L .
DRUG DISCOVERY TODAY, 2003, 8 (16) :755-761
[5]   Membrane protein microarrays [J].
Fang, Y ;
Frutos, AG ;
Lahiri, J .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (11) :2394-2395
[6]  
HAGA K, 1986, J BIOL CHEM, V261, P133
[7]  
HIRATA H, 1986, TECHNIQUES ANAL MEMB, P77
[8]   G protein-coupled receptors I. Diversity of receptor-ligand interactions [J].
Ji, TH ;
Grossmann, M ;
Ji, IH .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (28) :17299-17302
[9]   Flow-mediated on-surface reconstitution of G-protein coupled receptors for applications in surface plasmon resonance biosensors [J].
Karlsson, OP ;
Löfås, S .
ANALYTICAL BIOCHEMISTRY, 2002, 300 (02) :132-138
[10]  
Loidl-Stahlhofen A, 2001, ADV MATER, V13, P1829, DOI 10.1002/1521-4095(200112)13:23<1829::AID-ADMA1829>3.3.CO