Biacore analysis with stabilized G-protein-coupled receptors

被引：49

作者：

Rich, Rebecca L. ^{[1
]}

Errey, James ^{[2
]}

Marshall, Fiona ^{[2
]}

Myszka, David G. ^{[1
]}

机构：

[1] Univ Utah, Sch Med, Ctr Biomol Interact Anal, Salt Lake City, UT 84132 USA

[2] Heptares Therapeut, Welwyn Garden City AL7 3AX, Herts, England

来源：

ANALYTICAL BIOCHEMISTRY | 2011年 / 409卷 / 02期

基金：

美国国家卫生研究院;

关键词：

Surface plasmon resonance (SPR); Protein-protein; Kinetics; BIOSENSOR; THERMOSTABILIZATION; BINDING;

D O I：

10.1016/j.ab.2010.10.008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Using stabilized forms of beta(1) adrenergic and A(2A) adenosine G-protein-coupled receptors, we applied Biacore to monitor receptor activity and characterize binding constants of small-molecule antagonists spanning more than 20,000-fold in affinity. We also illustrate an improved method for tethering His-tagged receptors on NTA (carboxymethylated dextran preimmobilized with nitrilotriacetic acid) chips to yield stable, high-capacity, high-activity surfaces as well as a novel approach to regenerate receptor binding sites. Based on our success with this approach, we expect that the combination of stabilized receptors with biosensor technology will become a common method for characterizing members of this receptor family. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：267 / 272

页数：6

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