The neuronal Ca(2+) -binding protein 2 (NECAB2) interacts with the adenosine A(2A) receptor and modulates the cell surface expression and function of the receptor.
Heptaspanning membrane also known as G protein-coupled receptors (GPCR) do interact with a variety of intracellular proteins whose function is regulate receptor traffic and/or signaling. Using a yeast two-hybrid screen, NECAB2, a neuronal calcium binding protein, was identified as a binding partner for the adenosine A(2A) receptor (A(2A)R) interacting with its C-terminal domain. Co-localization, co-immunoprecipitation and pull-down experiments showed a close and specific interaction between A(2A)R and NECAB2 in both transfected HEK-293 cells and also in rat striatum. Immunoelectron microscopy detection of NECAB2 and A(2A)R in the rat striatopallidal structures indicated that both proteins are co-distributed in the same glutamatergic nerve terminals. The interaction of NECAB2 with A(2A)R modulated the cell surface expression, the ligand-dependent internalization and the receptor-mediated activation of the MAPK pathway. Overall, these results show that A(2A)R interacts with NECAB2 in striatal neurones co-expressing the two proteins and that the interaction is relevant for A(2A)R function.
Canela
Laia
L
Institut d'Investigacions Biomèdiques August Pi i Sunyer and Departament de Bioquímica i Biologia Molecular, Universitat de Barcelona, Barcelona, Spain.
Luján
Rafael
R
Lluís
Carme
C
Burgueño
Javier
J
Mallol
Josefa
J
Canela
Enric I
EI
Franco
Rafael
R
Ciruela
Francisco
F
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D003342:Corpus Striatum / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D004268:DNA-Binding Proteins; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D016253:Microscopy, Immunoelectron / Q000379:methods; D010627:Phenethylamines / Q000494:pharmacology; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D051381:Rats; D043683:Receptors, Adenosine A2 / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques
D000959:Antihypertensive Agents; D002135:Calcium-Binding Proteins; D004268:DNA-Binding Proteins; D010627:Phenethylamines; D043683:Receptors, Adenosine A2; C061282:2-(4-(2-carboxyethyl)phenethylamino)-5'-N-ethylcarboxamidoadenosine; D048049:Extracellular Signal-Regulated MAP Kinases; C085578:Ca-binding protein 2; D000241:Adenosine