Ligand binding analysis of soluble interleukin-2 receptor complexes by surface plasmon resonance.
Knowledge of the kinetic binding characteristics is often critical to the development of ligand/receptor structure-activity relationships. To better understand the contribution of each of the subunits to ligand binding in the multimeric interleukin-2 receptor system, we have previously prepared stable solution complexes of the alpha- and beta-subunits. In this study, we have employed surface plasmon resonance biosensor methodology (BIAcore) to evaluate both the kinetic and equilibrium binding constants for these complexes. The structural nature of the complexes facilitated immobilization on the sensor surfaces in a manner that minimized interference with ligand interactions. The interleukin-2 receptor complex surfaces displayed excellent binding capacity and stability toward regeneration. In all cases where the binding constants were measurable, the values determined for interleukin-2 were in good agreement with those previously determined by other methods. When interleukin-2 analogs with receptor subunit specific mutations were employed, the binding parameters were consistent with the nature of the mutations. The combination of coiled-coil-mediated solution assembly and surface plasmon resonance analysis of ligand binding provides a powerful approach to the study of multimeric cytokine receptor systems.
Wu
Z
Z
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Johnson
K W
KW
Choi
Y
Y
Ciardelli
T L
TL
D016477:Artifacts; D015374:Biosensing Techniques; D003627:Data Interpretation, Statistical; D005069:Evaluation Studies as Topic; D007376:Interleukin-2 / Q000031:analogs & derivatives / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D046911:Macromolecular Substances; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015375:Receptors, Interleukin-2 / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012995:Solubility; D013499:Surface Properties
D007376:Interleukin-2; D008024:Ligands; D046911:Macromolecular Substances; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins