{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T05:35:31Z","timestamp":1788413731809,"version":"build-2803163510"},"reference-count":50,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2025,10,18]],"date-time":"2025-10-18T00:00:00Z","timestamp":1760745600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["GM130389"],"award-info":[{"award-number":["GM130389"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003977","name":"Israel Science Foundation","doi-asserted-by":"publisher","award":["1706\/22"],"award-info":[{"award-number":["1706\/22"]}],"id":[{"id":"10.13039\/501100003977","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001698","name":"Israel Cancer Research Fund","doi-asserted-by":"publisher","award":["22\u2010203\u2010RCDA"],"award-info":[{"award-number":["22\u2010203\u2010RCDA"]}],"id":[{"id":"10.13039\/100001698","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2025,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The RI\u03b2 subunit of cAMP\u2010dependent protein kinase (PKA) is highly expressed in the brain, yet it remains the least studied of the PKA regulatory subunits (R). As pathologic variants of its gene are increasingly implicated in neurodevelopmental disorders, neurodegeneration, and cancer, gaining more information about the structure\/function of RI\u03b2, and how it differs from RI\u03b1, has become increasingly important. We previously reported the structure of the RI\u03b2\n                    <jats:sub>2<\/jats:sub>\n                    C\n                    <jats:sub>2<\/jats:sub>\n                    holoenzyme, which revealed a novel conformation where ATP binding was stabilized by a head\u2010to\u2010head anti\u2010parallel packing of the C\u2010tail wrapped around the N\u2010lobe of the catalytic subunit (C). Although visible, the Dimerization\/Docking Domain was poorly folded and reduced. Since RI\u03b2 is oxidized in brain tissues, we asked if oxidation or binding of an A Kinase Anchoring Protein (AKAP) would affect the holoenzyme structure. Oxidation or addition of an AKAP peptide to crystals led to the release of nucleotide. To capture this at higher resolution we crystallized RI\u03b2\n                    <jats:sub>2<\/jats:sub>\n                    C\n                    <jats:sub>2<\/jats:sub>\n                    in the presence of an AKAP peptide. This new structure represents an RI\u03b2:C heterodimer. Density for the D\/D domain was missing; ATP was absent, the kinase adopted an open conformation, and the C\u2010terminus of the RI\u03b2 subunit was no longer resolved. Because the crosstalk between ATP and cAMP in the R:C complex appears to be mediated by the two N3A motifs (N3A\n                    <jats:sup>A<\/jats:sup>\n                    and N3A\n                    <jats:sup>B<\/jats:sup>\n                    ) as well as by the linker, which in free RI\u03b2 is intrinsically disordered, we describe the conserved features of these two motifs as well as the linker and show how each contributes in a unique but coordinated way to allosteric activation of RI\u03b2 holoenzymes by cAMP. A key difference in our RI\u03b2:C structure is the rotation of the side chain of W260 at the N\u2010terminus of the \u03b1A Helix in N3A\n                    <jats:sup>B<\/jats:sup>\n                    . W260, at the R:C interface in the holoenzyme, is also the capping residue for cAMP bound to CNB\u2010A, so we may have actually captured the first step in cAMP activation.\n                  <\/jats:p>","DOI":"10.1002\/pro.70332","type":"journal-article","created":{"date-parts":[[2025,10,18]],"date-time":"2025-10-18T13:54:43Z","timestamp":1760795683000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["<scp>N3A<\/scp>\n                    motifs in\n                    <scp>RI\u03b2<\/scp>\n                    mediate allosteric crosstalk between\n                    <scp>cAMP<\/scp>\n                    and\n                    <scp>ATP<\/scp>\n                    in\n                    <scp>PKA<\/scp>\n                    activation"],"prefix":"10.1002","volume":"34","author":[{"given":"Jian","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Pharmacology University of California at San Diego  San Diego California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica G. H.","family":"Bruystens","sequence":"additional","affiliation":[{"name":"Department of Pharmacology University of California at San Diego  San Diego California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Puspashree","family":"Sahoo","sequence":"additional","affiliation":[{"name":"The Azrieli Faculty of Medicine Bar Ilan University  Safed Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9","family":"Bubis","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Universidad Sim\u00f3n Bol\u00edvar  Caracas Venezuela"},{"name":"Direcci\u00f3n de Salud, Fundaci\u00f3n Estudios Avanzados IDEA  Caracas Venezuela"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo A.","family":"Maillard","sequence":"additional","affiliation":[{"name":"Department of Chemistry Georgetown University  Washington, DC USA"},{"name":"Institute for Soft Matter Synthesis and Metrology Georgetown University  Washington, DC USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan S.","family":"Taylor","sequence":"additional","affiliation":[{"name":"Department of Pharmacology University of California at San Diego  San Diego California USA"},{"name":"Department of Biochemistry and Molecular Biophysics University of California at San Diego  San Diego California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7850-4143","authenticated-orcid":false,"given":"Ronit","family":"Ilouz","sequence":"additional","affiliation":[{"name":"The Azrieli Faculty of Medicine Bar Ilan University  Safed Israel"},{"name":"The Leslie &amp; Susan Goldschmied (Gonda) Multidisciplinary Brain Research Center Bar\u2010Ilan University  Ramat\u2010Gan Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,10,18]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1002305"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2013.03.010"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1608061113"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M003961200"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.70098"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awae154"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0408579102"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2010.12.005"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M114.568907"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M603952200"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.09.014"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2013.10.012"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)48122-7"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.13726"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.395970"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(89)90116-5"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi001563q"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00239-001-0013-1"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2019.03.001"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2023.104790"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)42419-8"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)42999-1"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-11930-2"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.00237.2019"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi982672w"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.12.8057"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1209538109"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.17681"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2007-8-12-r264"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.07.018"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1104607"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2006.09.015"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.45.28431"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1107\/S2059798319011471"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1906036116"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41436-021-01152-7"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1038\/290744a0"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/6663"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(97)76066-X"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2005.04.013"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)77178-6"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2009.12.012"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.7638597"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1002\/iub.2714"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm3432"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)77177-4"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)34204-8"},{"key":"e_1_2_9_49_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1146447"},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1213979"},{"key":"e_1_2_9_51_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444993000423"}],"container-title":["Protein 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