{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,9]],"date-time":"2026-05-09T01:00:19Z","timestamp":1778288419689,"version":"3.51.4"},"reference-count":52,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2024,1,5]],"date-time":"2024-01-05T00:00:00Z","timestamp":1704412800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000152","name":"Division of Molecular and Cellular Biosciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000152","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteins"],"published-print":{"date-parts":[[2024,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Human acyl protein thioesterases (APTs) catalyze the depalmitoylation of\n                    <jats:italic>S<\/jats:italic>\n                    \u2010acylated proteins attached to the plasma membrane, facilitating reversible cycles of membrane anchoring and detachment. We previously showed that a bacterial APT homologue, FTT258 from the gram\u2010negative pathogen\n                    <jats:italic>Francisella tularensis<\/jats:italic>\n                    , exists in equilibrium between a closed and open state based on the structural dynamics of a flexible loop overlapping its active site. Although the structural dynamics of this loop are not conserved in human APTs, the amino acid sequence of this loop is highly conserved, indicating essential but divergent functions for this loop in human APTs. Herein, we investigated the role of this loop in regulating the catalytic activity, ligand binding, and protein folding of human APT1, a depalmitoylase connected with cancer, immune, and neurological signaling. Using a combination of substitutional analysis with kinetic, structural, and biophysical characterization, we show that even in its divergent structural location in human APT1 that this loop still regulates the catalytic activity of APT1 through contributions to ligand binding and substrate positioning. We confirmed previously known roles for multiple residues (Phe72 and Ile74) in substrate binding and catalysis while adding new roles in substrate selectivity (Pro69), in catalytic stabilization (Asp73 and Ile75), and in transitioning between the membrane binding \u03b2\u2010tongue and substrate\u2010binding loops (Trp71). Even conservative substitution of this tryptophan (Trp71) fulcrum led to complete loss of catalytic activity, a 13\u00b0C decrease in total protein stability, and drastic drops in ligand affinity, indicating that the combination of the size, shape, and aromaticity of Trp71 are essential to the proper structure of APT1. Mixing buried hydrophobic surface area with contributions to an exposed secondary surface pocket, Trp71 represents a previously unidentified class of essential tryptophans within \u03b1\/\u03b2 hydrolase structure and a potential allosteric binding site within human APTs.\n                  <\/jats:p>","DOI":"10.1002\/prot.26661","type":"journal-article","created":{"date-parts":[[2024,1,5]],"date-time":"2024-01-05T05:38:54Z","timestamp":1704433134000},"page":"693-704","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A conserved but structurally divergent loop in acyl protein thioesterase 1 regulates its catalytic activity, ligand binding, and folded stability"],"prefix":"10.1002","volume":"92","author":[{"suffix":"III","given":"William Trey","family":"Harris","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry Butler University  Indianapolis Indiana USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isabelle","family":"Altieri","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Butler University  Indianapolis Indiana USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isabella","family":"Gieck","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Butler University  Indianapolis Indiana USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2694-3915","authenticated-orcid":false,"given":"R. Jeremy","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Butler University  Indianapolis Indiana USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,1,5]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.15781"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1080\/10409238.2017.1287161"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.phrs.2016.06.008"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbcan.2015.06.004"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.362"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20140235"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-021-00785-8"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-019-0399-y"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-02655-1"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.2262"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abj8633"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M202505200"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.6b00720"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsmedchemlett.2c00174"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2021.656440"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12964-019-0405-8"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2018.10.005"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-021-00753-2"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(00)00529-3"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.15071"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(97)00178-0"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/362814a0"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4089"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-38464-y"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.446625"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2015.09.006"},{"key":"e_1_2_11_28_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens11111245"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2018.05.012"},{"key":"e_1_2_11_30_1","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.M019851"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1080\/10409238.2018.1488804"},{"key":"e_1_2_11_32_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.7b00263"},{"key":"e_1_2_11_33_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA118.003972"},{"key":"e_1_2_11_34_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.6b01057"},{"key":"e_1_2_11_35_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi501108u"},{"key":"e_1_2_11_36_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed400783e"},{"key":"e_1_2_11_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201300085"},{"key":"e_1_2_11_38_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed500452f"},{"key":"e_1_2_11_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2012.05.009"},{"key":"e_1_2_11_40_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201700278"},{"key":"e_1_2_11_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.321"},{"key":"e_1_2_11_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2016.12.007"},{"key":"e_1_2_11_43_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsomega.3c00534"},{"key":"e_1_2_11_44_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.25285"},{"key":"e_1_2_11_45_1","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.M060673"},{"key":"e_1_2_11_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcatb.2014.08.008"},{"key":"e_1_2_11_47_1","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/gzp009"},{"key":"e_1_2_11_48_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2022.879251"},{"key":"e_1_2_11_49_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.113135"},{"key":"e_1_2_11_50_1","doi-asserted-by":"publisher","DOI":"10.1039\/C7SC02805A"},{"key":"e_1_2_11_51_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-020-0555-4"},{"key":"e_1_2_11_52_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg500"},{"key":"e_1_2_11_53_1","doi-asserted-by":"publisher","DOI":"10.1101\/gr.849004"}],"container-title":["Proteins: 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