Use of Kikume green-red fusions to study the influence of pharmacological chaperones on trafficking of G protein-coupled receptors.
In this study we demonstrate that the photoconvertible monomeric Kikume green-red (mKikGR) protein is suitable to study trafficking of G protein-coupled receptors. Taking mKikGR-tagged mutants of the vasopressin V(2) receptor (V(2)R) as models, we analyzed whether the V(2)R-specific pharmacological chaperone SR121463B influences receptor folding on a co- or post-translational level. Misfolded mKikGR-tagged V(2)Rs were completely photoconverted in the early secretory pathway yielding a red receptor population (already synthesized receptors) and an arising green receptor population (newly synthesized receptors). Trafficking of both receptor populations could be rescued by treatment with SR121463B demonstrating that the substance can act co- and post-translationally.
Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.
Ingrid
I
Ridelis
Antje
A
Schmidt
Anke
A
Teichmann
Jens
J
Furkert
Burkhard
B
Wiesner
Ralf
R
Schülein
D000595:Amino Acid Sequence; D065092:Antidiuretic Hormone Receptor Antagonists; D003116:Color; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009025:Morpholines / Q000737:chemistry / Q000378:metabolism; D017510:Protein Folding; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017483:Receptors, Vasopressin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013141:Spiro Compounds / Q000737:chemistry / Q000378:metabolism
D065092:Antidiuretic Hormone Receptor Antagonists; D008164:Luminescent Proteins; D018832:Molecular Chaperones; D009025:Morpholines; D043562:Receptors, G-Protein-Coupled; D017483:Receptors, Vasopressin; D011993:Recombinant Fusion Proteins; D013141:Spiro Compounds; C535245:mKikGR protein