{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T13:25:34Z","timestamp":1787059534666,"version":"build-2736575974"},"reference-count":41,"publisher":"American Chemical Society (ACS)","issue":"16","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016181","name":"San Jos? State University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100016181","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R16GM149377"],"award-info":[{"award-number":["R16GM149377"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R16GM150706"],"award-info":[{"award-number":["R16GM150706"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["S10GM154297"],"award-info":[{"award-number":["S10GM154297"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["1919817"],"award-info":[{"award-number":["1919817"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The NAD+-dependent deacetylase sirtuin-1 (SIRT1) is known to elicit cellular defenses against aging, cancer, and other aberrant pathologies. Previous studies have identified an intrinsically disordered region of SIRT1 comprised of N-terminal residues 1\u201352, herein referred to as motif A, which activates SIRT1 activity, likely through intramolecular interactions. Additionally, phosphorylation of N-terminal residues Ser27 and Ser47 has been shown to be important for regulating SIRT1 activity and stability. The lack of in vitro characterization of these effects hampers our further understanding of the role of motif A in SIRT1 regulation. In this study, we elucidate the role phosphorylation plays in motif A\u2019s structure as well as its regulatory effects on SIRT1 activity against Ac-p65. We find that a phosphomimetic mutation at Ser27 significantly increases the activation effect of motif A toward SIRT1. This result is supported by molecular dynamics simulations of the phosphomimetics, which reveal stabilization of different transient structures for motif A depending on whether Ser27 and Ser47 have been modified. A key finding suggested by this study is that phosphorylation of S27 appears to activate SIRT1 by causing motif A, which is intrinsically disordered in the WT, to become more rigid. This conclusion is based on both the experimental findings and simulation results. While more experiments are necessary to confirm the MD simulation results, these findings still bring us a step closer to understanding SIRT1 regulation, specifically the role played by phosphorylation within the N-terminal disordered region.<\/jats:p>","DOI":"10.1021\/acs.biochem.6c00333","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:14:41Z","timestamp":1785849281000},"page":"2485-2494","source":"Crossref","is-referenced-by-count":0,"title":["The Effects\nof Phosphorylation on the Structure and\nFunction of Motif A, an Intrinsically Disordered Region within SIRT1"],"prefix":"10.1021","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3066-3685","authenticated-orcid":true,"given":"Sabrina M.","family":"Richter","sequence":"first","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hoang-Long","family":"Bui","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Addison","family":"Chen","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chloe","family":"Tannous","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin R.","family":"Butler","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophia D.","family":"Bennett","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samson Quang-Anh","family":"Nguyen","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin","family":"Prado","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan","family":"Nhan","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayan","family":"Mohamed","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isaac A.","family":"DuBois","sequence":"additional","affiliation":[{"name":"Grand Valley State University , , , ,","place":["Allendale, Michigan, United States, 49401"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emile","family":"Tadros","sequence":"additional","affiliation":[{"name":"Grand Valley State University , , , ,","place":["Allendale, Michigan, United States, 49401"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nhu Tam","family":"Thai","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Selina","family":"Lima Guan","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5996-3790","authenticated-orcid":true,"given":"Carla Marie","family":"Peralta","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andy","family":"Kwong","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura M.L.","family":"Hawk","sequence":"additional","affiliation":[{"name":"Grand Valley State University , , , ,","place":["Allendale, Michigan, United States, 49401"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2559-5103","authenticated-orcid":true,"given":"Gianmarc","family":"Grazioli","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3545-5323","authenticated-orcid":true,"given":"Ningkun","family":"Wang","sequence":"additional","affiliation":[{"name":"San Jose\u0301 State University , , , ,","place":["San Jose\u0301, California, United States, 95112"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"key":"2026081808122786700_cit1","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1016\/S0092-8674(01)00527-X","article-title":"hSIR2SIRT1 functions\nas an NAD-dependent p53 deacetylase","volume":"107","author":"Vaziri","year":"2001","journal-title":"Cell"},{"key":"2026081808122786700_cit2","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1038\/nature03354","article-title":"Nutrient control\nof glucose homeostasis through a complex\nof PGC-1\u03b1 and SIRT1","volume":"434","author":"Rodgers","year":"2005","journal-title":"Nature"},{"key":"2026081808122786700_cit3","doi-asserted-by":"publisher","first-page":"2369","DOI":"10.1038\/sj.emboj.7600244","article-title":"Modulation of NF-\u03baB-dependent\ntranscription and cell survival by the SIRT1 deacetylase","volume":"23","author":"Yeung","year":"2004","journal-title":"EMBO J."},{"key":"2026081808122786700_cit4","doi-asserted-by":"publisher","first-page":"17023","DOI":"10.1038\/sigtrans.2017.23","article-title":"NF-\u03baB signaling\nin inflammation","volume":"2","author":"Liu","year":"2017","journal-title":"Signal Transduction Targeted\nTher."},{"key":"2026081808122786700_cit5","doi-asserted-by":"publisher","first-page":"6823","DOI":"10.2174\/1381612811319380014","article-title":"Acetylome\nregulation by sirtuins in the brain: from normal physiology to aging\nand pathology","volume":"19","author":"Michan","year":"2013","journal-title":"Curr. Pharm. Des."},{"key":"2026081808122786700_cit6","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1021\/jm301431y","article-title":"The 2.5\n\u00c5 Crystal Structure of the SIRT1 Catalytic\nDomain Bound to Nicotinamide Adenine Dinucleotide (NAD +) and an Indole\n(EX527 Analogue) Reveals a Novel Mechanism of Histone Deacetylase\nInhibition","volume":"56","author":"Zhao","year":"2013","journal-title":"J. Med. Chem."},{"key":"2026081808122786700_cit7","doi-asserted-by":"publisher","first-page":"1316","DOI":"10.1101\/gad.265462.115","article-title":"Structural basis for allosteric,\nsubstrate-dependent stimulation of SIRT1 activity by resveratrol","volume":"29","author":"Cao","year":"2015","journal-title":"Genes Dev."},{"key":"2026081808122786700_cit8","doi-asserted-by":"publisher","first-page":"7645","DOI":"10.1038\/ncomms8645","article-title":"Crystallographic\nstructure\nof a small molecule SIRT1 activator-enzyme complex","volume":"6","author":"Dai","year":"2015","journal-title":"Nat. Commun."},{"key":"2026081808122786700_cit9","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1006\/bbrc.2000.3000","article-title":"Phylogenetic\nClassification of Prokaryotic and Eukaryotic Sir2-like Proteins","volume":"273","author":"Frye","year":"2000","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2026081808122786700_cit10","doi-asserted-by":"publisher","first-page":"2468","DOI":"10.1074\/jbc.M111.285031","article-title":"SIRT1 Contains\nN- and C-terminal Regions That Potentiate Deacetylase Activity","volume":"287","author":"Pan","year":"2012","journal-title":"J. Biol. Chem."},{"key":"2026081808122786700_cit11","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1016\/j.molcel.2011.07.038","article-title":"Peptide Switch Is Essential for Sirt1 Deacetylase Activity","volume":"44","author":"Kang","year":"2011","journal-title":"Mol. Cell."},{"key":"2026081808122786700_cit12","doi-asserted-by":"publisher","first-page":"712","DOI":"10.1038\/nature06261","article-title":"Small molecule activators of SIRT1 as therapeutics\nfor the treatment of type 2 diabetes","volume":"450","author":"Milne","year":"2007","journal-title":"Nature"},{"key":"2026081808122786700_cit13","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1016\/j.molcel.2007.08.030","article-title":"Active\nRegulator\nof SIRT1 Cooperates with SIRT1 and Facilitates Suppression of p53\nActivity","volume":"28","author":"Kim","year":"2007","journal-title":"Mol. Cell."},{"key":"2026081808122786700_cit14","doi-asserted-by":"publisher","first-page":"985","DOI":"10.1016\/j.molcel.2018.10.007","article-title":"An Insulin-Responsive\nSensor in the SIRT1\nDisordered Region Binds DBC1 and PACS-2 to Control Enzyme Activity","volume":"72","author":"Krzysiak","year":"2018","journal-title":"Mol. Cell"},{"key":"2026081808122786700_cit15","doi-asserted-by":"publisher","first-page":"1665","DOI":"10.1016\/j.celrep.2015.02.036","article-title":"The N-Terminal\nDomain of SIRT1 Is a Positive Regulator\nof Endogenous SIRT1-Dependent Deacetylation and Transcriptional Outputs","volume":"10","author":"Ghisays","year":"2015","journal-title":"Cell Rep."},{"key":"2026081808122786700_cit16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0004020","article-title":"Phosphorylation\nRegulates SIRT1 Function","volume":"3","author":"Sasaki","year":"2008","journal-title":"PLoS One"},{"key":"2026081808122786700_cit17","doi-asserted-by":"publisher","first-page":"29","DOI":"10.3389\/fphar.2012.00029","article-title":"Regulation of Sirtuin Function by\nPosttranslational\nModifications","volume":"3","author":"Flick","year":"2012","journal-title":"Front. Pharmacol."},{"key":"2026081808122786700_cit18","doi-asserted-by":"publisher","first-page":"19100","DOI":"10.1074\/jbc.M111.240598","article-title":"Cancer Cell Survival\nFollowing DNA Damage-mediated Premature Senescence Is Regulated by\nMammalian Target of Rapamycin (mTOR)-dependent Inhibition of Sirtuin\n1","volume":"286","author":"Back","year":"2011","journal-title":"J. Biol. Chem."},{"key":"2026081808122786700_cit19","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1161\/CIRCULATIONAHA.112.118778","article-title":"Cyclin-Dependent Kinase 5\u2013Mediated\nHyperphosphorylation of Sirtuin-1 Contributes to the Development of\nEndothelial Senescence and Atherosclerosis","volume":"126","author":"Bai","year":"2012","journal-title":"Circulation"},{"key":"2026081808122786700_cit20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0008414","article-title":"JNK1 Phosphorylates\nSIRT1 and Promotes Its Enzymatic Activity","volume":"4","author":"Nasrin","year":"2009","journal-title":"PLoS\nOne"},{"key":"2026081808122786700_cit21","doi-asserted-by":"publisher","first-page":"3091","DOI":"10.4161\/cc.7.19.6799","article-title":"JNK2-dependent\nregulation of SIRT1 protein stability","volume":"7","author":"Ford","year":"2008","journal-title":"Cell\nCycle"},{"key":"2026081808122786700_cit22","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1309354110","article-title":"Ca 2+ \/calmodulin-dependent\nprotein kinase kinase \u03b2 phosphorylation of Sirtuin 1 in endothelium\nis atheroprotective","volume":"110","author":"Wen","year":"2013","journal-title":"Proc. Natl. Acad. Sci."},{"key":"2026081808122786700_cit23","doi-asserted-by":"publisher","first-page":"478","DOI":"10.1074\/jbc.M807017200","article-title":"Six Classes\nof Nuclear\nLocalization Signals Specific to Different Binding Grooves of Importin\n\u03b1*","volume":"284","author":"Kosugi","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2026081808122786700_cit24","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1016\/j.ab.2009.07.019","article-title":"A continuous microplate assay for\nsirtuins and nicotinamide-producing enzymes","volume":"394","author":"Smith","year":"2009","journal-title":"Anal. Biochem."},{"key":"2026081808122786700_cit25","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate\nstructure prediction of biomolecular\ninteractions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"2026081808122786700_cit26","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1021\/acs.jctc.5b00935","article-title":"CHARMM-GUI\nInput Generator for NAMD, GROMACS, AMBER,\nOpenMM, and CHARMM\/OpenMM Simulations Using the CHARMM36 Additive\nForce Field","volume":"12","author":"Lee","year":"2016","journal-title":"J. Chem. Theory Comput."},{"key":"2026081808122786700_cit27","doi-asserted-by":"publisher","first-page":"1859","DOI":"10.1002\/jcc.20945","article-title":"CHARMM-GUI: A web-based\ngraphical\nuser interface for CHARMM","volume":"29","author":"Jo","year":"2008","journal-title":"J. Comput. Chem."},{"key":"2026081808122786700_cit28","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1021\/ja00392a016","article-title":"Quantum\nand statistical mechanical studies of liquids. 10. Transferable intermolecular\npotential functions for water, alcohols, and ethers. Application to\nliquid water","volume":"103","author":"Jorgensen","year":"1981","journal-title":"J. Am. Chem. Soc."},{"key":"2026081808122786700_cit29","doi-asserted-by":"publisher","first-page":"1781","DOI":"10.1002\/jcc.20289","article-title":"Scalable molecular dynamics with\nNAMD","volume":"26","author":"Phillips","year":"2005","journal-title":"J. Comput. Chem."},{"key":"2026081808122786700_cit30","doi-asserted-by":"publisher","first-page":"1545","DOI":"10.1002\/jcc.21287","article-title":"CHARMM: the biomolecular\nsimulation program","volume":"30","author":"Brooks","year":"2009","journal-title":"J. Comput. Chem."},{"key":"2026081808122786700_cit31","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1038\/nmeth.4067","article-title":"CHARMM36m:\nan improved force field for folded and intrinsically disordered proteins","volume":"14","author":"Huang","year":"2017","journal-title":"Nat. Methods"},{"key":"2026081808122786700_cit32","volume-title":"MDAnalysis: A Python Package for the Rapid Analysis of Molecular\nDynamics Simulations","author":"Gowers","year":"2016"},{"key":"2026081808122786700_cit33","doi-asserted-by":"publisher","first-page":"2319","DOI":"10.1002\/jcc.21787","article-title":"MDAnalysis:\nA toolkit for the analysis of molecular dynamics simulations","volume":"32","author":"Michaud-Agrawal","year":"2011","journal-title":"J. Comput. Chem."},{"key":"2026081808122786700_cit34","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/0263-7855(96)00018-5","article-title":"VMD: Visual\nmolecular dynamics","volume":"14","author":"Humphrey","year":"1996","journal-title":"J. Mol. Graph"},{"key":"2026081808122786700_cit35","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1038\/s41586-020-2649-2","article-title":"Array programming with NumPy","volume":"585","author":"Harris","year":"2020","journal-title":"Nature"},{"key":"2026081808122786700_cit36","doi-asserted-by":"publisher","first-page":"90","DOI":"10.1109\/MCSE.2007.55","article-title":"Matplotlib:\nA 2D Graphics Environment","volume":"9","author":"Hunter","year":"2007","journal-title":"Comput. Sci. Eng."},{"key":"2026081808122786700_cit37","volume-title":"The Pandas Development Teampandas-dev\/pandas","author":"Zenodo.","year":"2026"},{"key":"2026081808122786700_cit38","doi-asserted-by":"publisher","first-page":"W176","DOI":"10.1093\/nar\/gkae385","article-title":"AIUPred: combining energy estimation\nwith deep learning\nfor the enhanced prediction of protein disorder","volume":"52","author":"Erdo\u030bs","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"2026081808122786700_cit39","doi-asserted-by":"publisher","first-page":"2745","DOI":"10.1093\/bioinformatics\/btp518","article-title":"ANCHOR: web server\nfor predicting protein binding regions in disordered proteins","volume":"25","author":"Doszta\u0301nyi","year":"2009","journal-title":"Bioinforma. Oxf. Engl."},{"key":"2026081808122786700_cit40","doi-asserted-by":"publisher","first-page":"i75","DOI":"10.1093\/bioinformatics\/bts209","article-title":"MoRFpred, a computational\ntool for sequence-based prediction and characterization of short disorder-to-order\ntransitioning binding regions in proteins","volume":"28","author":"Disfani","year":"2012","journal-title":"Bioinformatics"},{"key":"2026081808122786700_cit41","doi-asserted-by":"publisher","first-page":"566","DOI":"10.1002\/prot.340230412","article-title":"Knowledge-based protein\nsecondary structure assignment","volume":"23","author":"Frishman","year":"1995","journal-title":"Proteins: struct.,\nFunct., Bioinf."}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.biochem.6c00333","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/16\/2485\/66320974\/acs.biochem.6c00333.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/16\/2485\/66320974\/acs.biochem.6c00333.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T12:38:23Z","timestamp":1787056703000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/65\/16\/2485\/5236212\/The-Effects-of-Phosphorylation-on-the-Structure"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,28]]},"references-count":41,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,7,28]]},"published-print":{"date-parts":[[2026,8,18]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.biochem.6c00333","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,28]]}}}