{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,6]],"date-time":"2025-11-06T11:52:01Z","timestamp":1762429921245},"reference-count":43,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2004,6,4]],"date-time":"2004-06-04T00:00:00Z","timestamp":1086307200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Neurochemistry"],"published-print":{"date-parts":[[2004,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Effects of protein kinase C on protein stability and activity of rat AANAT were investigated <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic>. When COS\u20107 cells transfected with AANAT cDNA were treated with phorbol 12\u2010myristate 13\u2010acetate (PMA), both the activity and protein level of AANAT were increased. These effects of PMA were blocked by GF109203X, a specific inhibitor of PKC. Moreover, PMA increased the phosphorylation of AANAT and induced the formation of AANAT\/14\u20103\u20103\u03b6 complex. PMA did not affect the basal level of cAMP and did not involve the potentiation of the cAMP production by forskolin, indicating that PKC\u2010dependent activation of adenylyl cyclase was excluded in transfected COS\u20107 cells. To identify which amino acids were phosphorylated by PKC, several conserved Thr and Ser residues in AANAT were targeted for site\u2010directed mutagenesis. Mutations of Thr29 and Ser203 prevented the increase of enzymatic activity and protein level mediated by PMA. To explore the nature of AANAT phosphorylation, purified rat AANAT was subjected to <jats:italic>in vitro<\/jats:italic> PKC kinase assay. PKC directly phosphorylated the rat recombinant AANAT. The phosphopeptides identified by mass spectrometric analysis, and western blotting indicated that Thr29 was one of target sites for PKC. To confirm the effects of the physiological activation of PKC, rat pineal glands were treated with \u03b1<jats:sub>1<\/jats:sub>\u2010adrenergic specific agonist phenylephrine. Phenylephrine caused the phosphorylation of endogenous AANAT whereas GF109203X or prazosin, an \u03b1<jats:sub>1<\/jats:sub>\u2010adrenergic\u2010specific antagonist, markedly inhibited it. These results suggest that AANAT was phosphorylated at Thr29 by PKC activation through the \u03b1<jats:sub>1<\/jats:sub>\u2010adrenergic receptor in rat pineal glands, and that its phosphorylation might contribute to the stability and the activity of AANAT.<\/jats:p>","DOI":"10.1111\/j.1471-4159.2004.02495.x","type":"journal-article","created":{"date-parts":[[2004,7,2]],"date-time":"2004-07-02T06:12:45Z","timestamp":1088748765000},"page":"442-454","source":"Crossref","is-referenced-by-count":31,"title":["Protein kinase C regulates the activity and stability of serotonin <i>N<\/i>\u2010acetyltransferase"],"prefix":"10.1111","volume":"90","author":[{"given":"Bo\u2010Hwa","family":"Choi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hee\u2010Don","family":"Chae","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tae\u2010Ju","family":"Park","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jisun","family":"Oh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinkyu","family":"Lim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shin\u2010Sung","family":"Kang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hyunjung","family":"Ha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kyong\u2010Tai","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,6,7]]},"reference":[{"key":"e_1_2_28_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.1.304"},{"key":"e_1_2_28_3_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.56.070187.001111"},{"key":"e_1_2_28_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/378783a0"},{"key":"e_1_2_28_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_2_28_6_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj1210561"},{"key":"e_1_2_28_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0197-0186(99)00086-8"},{"key":"e_1_2_28_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M005671200"},{"key":"e_1_2_28_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.270.5242.1681"},{"key":"e_1_2_28_10_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M011298200"},{"key":"e_1_2_28_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.1972.tb01458.x"},{"key":"e_1_2_28_12_1","doi-asserted-by":"crossref","first-page":"1804","DOI":"10.1210\/endo.142.5.8129","article-title":"Regulation of arylalkylamine N\u2010acetyltransferase\u20102 (AANAT2, EC 2.3.1.87) in the fish pineal organ: evidence for a role of proteasomal proteolysis","volume":"142","author":"Falcon J.","year":"2001","journal-title":"Endocrinology"},{"key":"e_1_2_28_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1476-5381.1994.tb13037.x"},{"key":"e_1_2_28_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-002-8517-4"},{"key":"e_1_2_28_15_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.141118798"},{"key":"e_1_2_28_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-002-0579-y"},{"key":"e_1_2_28_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.279.5355.1358"},{"key":"e_1_2_28_18_1","doi-asserted-by":"crossref","first-page":"9292","DOI":"10.1016\/S0021-9258(19)76538-7","article-title":"Protein kinase C: subcellular redistribution by increased Ca2+ influx: evidence that Ca2+\u2010dependent subcellular redistribution of protein kinase C is involved in potentiation of beta\u2010adrenergic stimulation of pineal cAMP and cGMP by K+ and A23187","volume":"263","author":"Ho A. 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