The 5-hydroxytryptamine(4a) receptor is palmitoylated at two different sites, and acylation is critically involved in regulation of receptor constitutive activity. Export

@article{citeulike:742126, abstract = {We have reported recently that the mouse 5-hydroxytryptamine(4a) (5-HT(4(a))) receptor undergoes dynamic palmitoylation (Ponimaskin, E. G., Schmidt, M. F., Heine, M., Bickmeyer, U., and Richter, D. W. (2001) Biochem. J. 353, 627-663). In the present study, conserved cysteine residues 328/329 in the carboxyl terminus of the 5-HT(4(a)) receptor were identified as potential acylation sites. In contrast to other palmitoylated G-protein-coupled receptors, the additional cysteine residue 386 positioned close to the COOH-terminal end of the receptor was also found to be palmitoylated. Using pulse and pulse-chase labeling techniques, we demonstrated that palmitoylation of individual cysteines is a reversible process and that agonist stimulation of the 5-HT(4(a)) receptor independently increases the rate of palmitate turnover for both acylation sites. Analysis of acylation-deficient mutants revealed that non-palmitoylated 5-HT(4(a)) receptors were indistinguishable from the wild type in their ability to interact with G(s), to stimulate the adenylyl cyclase activity and to activate cyclic nucleotide-sensitive cation channels after agonist stimulation. The most distinctive finding of the present study was the ability of palmitoylation to modulate the agonist-independent constitutive 5-HT(4(a)) receptor activity. We demonstrated that mutation of the proximal palmitoylation site (Cys(328) --> Ser/Cys(329) --> Ser) significantly increases the capacity of receptors to convert from the inactive (R) to the active (R*) form in the absence of agonist. In contrast, the rate of isomerization from R to R* for the Cys(386) --> Ser as well as for the triple, non-palmitoylated mutant (Cys(328) --> Ser/Cys(329) --> Ser/Cys(386) -->Ser) was similar to that obtained for the wild type.}, address = {Abteilung Neuro- und Sinnesphysiologie, Physiologisches Institut, Universit\"{a}t G\"{o}ttingen, Humboldtallee 23, D-37073 G\"{o}ttingen, Germany.}, author = {Ponimaskin, E. G. and Heine, M. and Joubert, L. and Sebben, M. and Bickmeyer, U. and Richter, D. W. and Dumuis, A.}, citeulike-article-id = {742126}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M106529200}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/11706023}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=11706023}, day = {25}, doi = {10.1074/jbc.M106529200}, issn = {0021-9258}, journal = {J Biol Chem}, keywords = {5-ht4, acylation, palmitoylation}, month = {January}, number = {4}, pages = {2534--2546}, posted-at = {2006-07-07 23:51:35}, priority = {2}, title = {The 5-hydroxytryptamine(4a) receptor is palmitoylated at two different sites, and acylation is critically involved in regulation of receptor constitutive activity.}, url = {http://dx.doi.org/10.1074/jbc.M106529200}, volume = {277}, year = {2002} }

TY - JOUR ID - citeulike:742126 L3 - citeulike-article-id:742126 N2 - We have reported recently that the mouse 5-hydroxytryptamine(4a) (5-HT(4(a))) receptor undergoes dynamic palmitoylation (Ponimaskin, E. G., Schmidt, M. F., Heine, M., Bickmeyer, U., and Richter, D. W. (2001) Biochem. J. 353, 627-663). In the present study, conserved cysteine residues 328/329 in the carboxyl terminus of the 5-HT(4(a)) receptor were identified as potential acylation sites. In contrast to other palmitoylated G-protein-coupled receptors, the additional cysteine residue 386 positioned close to the COOH-terminal end of the receptor was also found to be palmitoylated. Using pulse and pulse-chase labeling techniques, we demonstrated that palmitoylation of individual cysteines is a reversible process and that agonist stimulation of the 5-HT(4(a)) receptor independently increases the rate of palmitate turnover for both acylation sites. Analysis of acylation-deficient mutants revealed that non-palmitoylated 5-HT(4(a)) receptors were indistinguishable from the wild type in their ability to interact with G(s), to stimulate the adenylyl cyclase activity and to activate cyclic nucleotide-sensitive cation channels after agonist stimulation. The most distinctive finding of the present study was the ability of palmitoylation to modulate the agonist-independent constitutive 5-HT(4(a)) receptor activity. We demonstrated that mutation of the proximal palmitoylation site (Cys(328) --> Ser/Cys(329) --> Ser) significantly increases the capacity of receptors to convert from the inactive (R) to the active (R*) form in the absence of agonist. In contrast, the rate of isomerization from R to R* for the Cys(386) --> Ser as well as for the triple, non-palmitoylated mutant (Cys(328) --> Ser/Cys(329) --> Ser/Cys(386) -->Ser) was similar to that obtained for the wild type. IS - 4 JF - J Biol Chem SN - 0021-9258 AD - Abteilung Neuro- und Sinnesphysiologie, Physiologisches Institut, Universität Göttingen, Humboldtallee 23, D-37073 Göttingen, Germany. EP - 2546 TI - The 5-hydroxytryptamine(4a) receptor is palmitoylated at two different sites, and acylation is critically involved in regulation of receptor constitutive activity. VL - 277 SP - 2534 KW - 5-ht4 KW - acylation KW - palmitoylation AU - Ponimaskin, EG AU - Heine, M AU - Joubert, L AU - Sebben, M AU - Bickmeyer, U AU - Richter, DW AU - Dumuis, A PY - 2002/01/25/ UR - http://dx.doi.org/10.1074/jbc.M106529200 ER -