G protein-coupled receptor kinase 3 (GRK3) in olfaction

Abstract

Like in other sensory systems, adaptation is an essential process in the olfactory system, required for its proper functioning. However, the precise molecular mechanism underlying the adaptation process has not been fully understood, especially at the receptor level. Here, we describe methods to evaluate the role of GRK3, one of the members of the GRK family responsible for the desensitization of non-olfactory G-protein-coupled receptor (GPCR), in desensitization of olfactory receptor (OR) using a heterologous expression system. As a parameter to characterize the degree of desensitization, we measure (1) the maximal response to an agonist by either cAMP or Ca 2+ imaging assay and (2) the kinetic time course for recovery to basal levels by Ca 2+ imaging assay. Differences in the degree of desensitization in the presence or absence of GRK3 can be examined by comparing these parameters, leading to evaluation of GRK3.