1Unit of Molecular Pharmacology and Pharmacognosy, Institute for Scientific Research and High Technology Services, Bld. 219, City of Knowledge, Clayton, Panama Republic
2Department of Developmental and Cell Biology, University of California, Irvine, California, USA
3Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California, USA
4Institut für Biochemie und Molekularbiologie, Nussallee 11, 53115, Bonn, Germany
5Department of Biotechnology, Acharya Nagarjuna University, Guntur, India
*For correspondence – c.caballerogeorge@gmail.com
Online published on 29 April, 2013.
Fluorescence spectroscopy is the most common non-radioactive technique used to study GPCR interactions with their ligands. Raster image correlation spectroscopy (RICS) exploits spatio-temporal correlation functions rather than the simple temporal correlations of conventional fluorescence correlation spectroscopy. In this paper we describe the use of RICS and the number and brightness method to determine the diffusion of a construct of endothelin ETA receptor with EGFP and the aggregation state in the cytoplasm. Our construct seems to locate mainly in the cytoplasm where it undergoes diffusion and it appears to be monomeric. Although our construct could not fully represent the native protein, we believe that the methodology we describe in this paper could be used by anyone in this field.
Endothelin ETA receptor, GPCR, live cell, imaging, RICS
