Membrane proteins, such as G protein-coupled receptors (GPCRs) and ion channels, represent important but technically challenging targets for the management of pain and other diseases. Studying their interactions has enabled the development of new therapeutics, diagnostics, and research reagents, but biophysical manipulation of membrane proteins is often difficult because of the requirement of most membrane proteins for an intact lipid bilayer. Here, we describe the use of virus-like particles as presentation vehicles for cellular membrane proteins (“Lipoparticles”). The methods for using Lipoparticles on optical biosensors, such as the BioRad ProteOn XPR36, are discussed as a means to characterize the kinetics, affinity, and specificity of antibody interactions using surface plasmon resonance detection.
Biosensor Virus-like particle Membrane protein GPCR
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We thank Sharon Willis for her help with Lipoparticle production and biosensor optimization and Laura Moriarty (BioRad) and Mohammed Yousef (BioRad) for their helpful discussions.
Navratilova I, Dioszegi M, Myszka DG (2006) Analyzing ligand and small molecule binding activity of solubilized GPCRs using biosensor technology. Anal Biochem 355(1):132–139PubMedCrossRefGoogle Scholar
Navratilova I, Sodroski J, Myszka DG (2005) Solubilization, stabilization, and purification of chemokine receptors using biosensor technology. Anal Biochem 339(2):271–281PubMedCrossRefGoogle Scholar
Rice PJ et al (2002) Human monocyte scavenger receptors are pattern recognition receptors for (1->3)-beta-D-glucans. J Leukoc Biol 72(1):140–146PubMedGoogle Scholar
Stenlund P, Babcock GJ, Sodroski J, Myszka DG (2003) Capture and reconstitution of G protein-coupled receptors on a biosensor surface. Anal Biochem 316(2):243–250PubMedCrossRefGoogle Scholar
Mobini R et al (2000) A monoclonal antibody directed against an autoimmune epitope on the human beta1-adrenergic receptor recognized in idiopathic dilated cardiomyopathy. . Hybridoma 19(2):135-142 (in eng)PubMedCrossRefGoogle Scholar
Peter JC, Eftekhari P, Billiald P, Wallukat G, Hoebeke J (2003) scFv single chain antibody variable fragment as inverse agonist of the beta2-adrenergic receptor. (Translated from eng). J Biol Chem 278(38):36740-36747 (in eng)PubMedCrossRefGoogle Scholar
Day PW et al (2007) A monoclonal antibody for G protein-coupled receptor crystallography. Nat Methods 4(11):927–929PubMedCrossRefGoogle Scholar
Tummino PJ, Copeland RA (2008) Residence time of receptor-ligand complexes and its effect on biological function. (Translated from eng). Biochemistry 47(20):5481-5492 (in eng)PubMedCrossRefGoogle Scholar
Canziani G et al (1999) Exploring biomolecular recognition using optical biosensors. (Translated from eng). Methods 19(2):253-269 (in eng)PubMedCrossRefGoogle Scholar
Willis S et al (2008) Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry 47(27):6988–6990PubMedCrossRefGoogle Scholar
Bravman T et al (2006) Exploring “one-shot” kinetics and small molecule analysis using the ProteOn XPR36 array biosensor. (Translated from eng). Anal Biochem 358(2):281-288 (in eng)PubMedCrossRefGoogle Scholar
Hoffman TL, Canziani G, Jia L, Rucker J, Doms RW (2000) A biosensor assay for studying ligand-membrane receptor interactions: binding of antibodies and HIV-1 Env to chemokine receptors. (Translated from eng). Proc Natl Acad Sci USA 97(21):11215-11220 (in eng)PubMedCrossRefGoogle Scholar
Karlsson R, Katsamba PS, Nordin H, Pol E, Myszka DG (2006) Analyzing a kinetic titration series using affinity biosensors. (Translated from eng). Anal Biochem 349(1):136-147 (in eng)PubMedCrossRefGoogle Scholar
Green RJ, Davies MC, Roberts CJ, Tendler SJ (1998) A surface plasmon resonance study of albumin adsorption to PEO-PPO-PEO triblock copolymers. (Translated from eng). J Biomed Mater Res 42(2):165-171 (in eng)PubMedCrossRefGoogle Scholar
Segura MM et al (2008) Identification of host proteins associated with retroviral vector particles by proteomic analysis of highly purified vector preparations. J Virol 82(3):1107–1117PubMedCrossRefGoogle Scholar
Myszka DG, He X, Dembo M, Morton TA, Goldstein B (1998) Extending the range of rate constants available from BIACORE: interpreting mass transport-influenced binding data. (Translated from eng). Biophys J 75(2):583-594 (in eng)PubMedCrossRefGoogle Scholar