Morphological and biochemical strategies for monitoring trafficking of epitope-tagged G protein-coupled receptors in agonist-naive and agonist-occupied states.
Epitope tagged alpha 2-AR subtypes have been used to address a variety of cell biological questions, and the strategies used are readily applicable to all GPCR as well as other cell surface proteins. We have provided detailed protocols for successful utilization of the epitope-tagged receptor in the studies of protein localization and trafficking in epithelial cells, and the mechanisms by which this is achieved. We have also described reversible biotinytion strategies to examine agonist-dependent (and independent) receptor turnover at the cell surface.
Wozniak
Magdalena
M
Renal Division, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Saunders
Christine
C
Schramm
Nicole
N
Keefer
Jeffrey R
JR
Limbird
Lee E
LE
D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin / Q000378:metabolism; D002455:Cell Division; D018604:Epitope Mapping; D019204:GTP-Binding Proteins / Q000378:metabolism*; D021381:Protein Transport; D011956:Receptors, Cell Surface / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*
D011956:Receptors, Cell Surface; D001710:Biotin; D019204:GTP-Binding Proteins