Abstract
Caveolin-1 (Cav1), is a structural protein component of many mammalian cell plasma membrane and is known to be involved in lipid and protein sorting, receptor desensitization, receptor trafficking, cell migration and many other cellular events. Here we determine if stable expression of Cav1 in cells alters the receptor organization prototype on the membrane. We use two different cell lines for this study: Fisher Rat Thyroid (FRTwt) cells that do not express detectable level of Cav1 and a sister line that is stably transfected with canine Cav1 protein (FRTcav). We express μ opioid receptors (MOR) tagged with either YFP (MOR-YFP) or CFP (MOR-CFP) in cells for different experiments. Forster resonance energy transfer (FRET) measurement between MOR-CFP and Gαi-YFP in FRTwt and FRTcav cells shows receptor sequestration in the presence of Cav1. We find that diffusion of MOR-YFP in plasma membrane of FRTcav cells is slower compared to FRTwt cells by scanning fluorescence correlation spectroscopy (scanning-FCS) experiments. Photon counting histogram (PCH) analyses provide higher average brightness for MOR-YFP in FRTcav cells. Taken together these data provide evidence for caveolin-assisted enhanced clustering of G-protein coupled receptors on the plasma membrane.
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