Characterization of functional roles of DRY motif in the 2nd intracellular loop of dopamine D2 and D3 receptors.
Dopamine D(2)R and D(3)R (D(2)R, D(3)R) show very high sequence homology and employ virtually identical signaling pathways even though D(2)R is 2 approximately 5 times more active. Among the structural motifs identified, a triplet sequence, Asp-Arg-Tyr (DRY motif), plays critical roles in the determination of receptor conformations for signaling and intracellular trafficking of G protein-coupled receptors by forming intramolecular interactions. Thus, it is possible that different signaling efficiencies of D(2)R and D(3)R might be caused by the receptor activation levels stabilized by their own DRY motifs. In this study, the Arg and Asp residues of D(2)R and D(3)R were mutated, and resulting changes in their signaling and intracellular trafficking properties were comparatively studied. Mutation of the Arg residues of D(2)R and D(3)R abolished their signaling but differently affected their intracellular localizations. The wildtype and R132H-D(2)R were expressed mainly on the plasma membrane. On the other hand, compared with the wildtype D(3)R, a substantial amount of R128H-D(3)R was localized intracellularly. The expression of receptor proteins on the plasma membrane and their signaling efficiencies were more drastically affected by the mutation of the Asp residue of D(3)R than D(2)R. Therefore, it was concluded that the different levels of conformational strain exerted by the DRY motif might partly determine the quantitative differences in the signaling efficiencies between D(2)R and D(3)R.
Kim
Ju-Heon
JH
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA.
Cho
Eun-Young
EY
Min
Chengchun
C
Park
Jae H
JH
Kim
Kyeong-Man
KM
D020816:Amino Acid Motifs; D001058:Apomorphine / Q000031:analogs & derivatives / Q000494:pharmacology; D001120:Arginine / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D018491:Dopamine Agonists / Q000494:pharmacology; D018492:Dopamine Antagonists / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D009154:Mutation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D019257:Quinpirole / Q000494:pharmacology; D017448:Receptors, Dopamine D2 / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D050637:Receptors, Dopamine D3 / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013134:Spiperone / Q000494:pharmacology; D013469:Sulpiride / Q000494:pharmacology; D014162:Transfection
D018491:Dopamine Agonists; D018492:Dopamine Antagonists; D017448:Receptors, Dopamine D2; D050637:Receptors, Dopamine D3; D019257:Quinpirole; D001224:Aspartic Acid; D013134:Spiperone; C009074:N-n-propylnorapomorphine; D013469:Sulpiride; D001120:Arginine; D000242:Cyclic AMP; D001058:Apomorphine