Quantitative approach for detection and characterization of formyl peptide receptor in solution using a photoaffinity ligand.

Abstract

In this paper we describe an assay method for the rapid detection and quantitation of human neutrophil FMLP (N-formyl-methionyl-leucyl-phenylalanine) receptor molecules. Although different assay methods are available to detect the receptor, none are rapid and at the same time allow quantitative detection of the binding affinity of the receptor molecule in solution. In our modified method, following a binding reaction using photoaffinity ligand, the amount of labeled ligand bound to the receptor is separated from the unbound one, thereby determining the binding affinity of the receptor protein. This simple procedure not only makes it possible to detect the recombinant FMLP receptor protein very rapidly, but also provides quantitative assessment of binding. This technique therefore allows a partial characterization (identification, determination of size and assessment of binding affinity) of receptor molecule in solution in less than 24 hours.