{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T17:31:37Z","timestamp":1770226297491,"version":"3.49.0"},"reference-count":79,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2021,3,19]],"date-time":"2021-03-19T00:00:00Z","timestamp":1616112000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemPhysChem"],"published-print":{"date-parts":[[2021,4,19]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Nitrilase 2 (Nit2) is a representative member of the nitrilase superfamily that catalyzes the hydrolysis of \u03b1\u2010ketosuccinamate into oxaloacetate. It has been associated with the metabolism of rapidly dividing cells like cancer cells. The catalytic mechanism of Nit2 employs a catalytic triad formed by Cys191, Glu81 and Lys150. The Cys191 and Glu81 play an active role during the catalytic process while the Lys150 is shown to play only a secondary role. The results demonstrate that the catalytic mechanism of Nit2 involves four steps. The nucleophilic attack of Cys191 to the \u03b1\u2010ketosuccinamate, the formation of two tetrahedral enzyme adducts and the hydrolysis of a thioacyl\u2010enzyme intermediate, from which results the formation of oxaloacetate and enzymatic turnover. The rate limiting step of the catalytic process is the formation of the first tetrahedral intermediate with a calculated activation free energy of 18.4\u2005kcal\/mol, which agrees very well with the experimental <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> (17.67\u2005kcal\/mol).<\/jats:p>","DOI":"10.1002\/cphc.202000751","type":"journal-article","created":{"date-parts":[[2021,1,19]],"date-time":"2021-01-19T08:08:50Z","timestamp":1611043730000},"page":"796-804","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["An Unsual Cys\u2010Glu\u2010Lys Catalytic Triad is Responsible for the Catalytic Mechanism of the Nitrilase Superfamily: A QM\/MM Study on Nit2"],"prefix":"10.1002","volume":"22","author":[{"given":"Carla S.","family":"Silva Teixeira","sequence":"first","affiliation":[{"name":"UCIBIO@REQUIMTE BioSIM Departamento de Biomedicina Faculdade de Medicina Universidade do Porto  Porto 4200-319 Portugal"}]},{"given":"S\u00e9rgio F.","family":"Sousa","sequence":"additional","affiliation":[{"name":"UCIBIO@REQUIMTE BioSIM Departamento de Biomedicina Faculdade de Medicina Universidade do Porto  Porto 4200-319 Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0342-7424","authenticated-orcid":false,"given":"Nuno M. F. S. A.","family":"Cerqueira","sequence":"additional","affiliation":[{"name":"UCIBIO@REQUIMTE BioSIM Departamento de Biomedicina Faculdade de Medicina Universidade do Porto  Porto 4200-319 Portugal"}]}],"member":"311","published-online":{"date-parts":[[2021,3,19]]},"reference":[{"key":"e_1_2_7_1_1","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2001-2-1-reviews0001"},{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0959-440X(02)00387-1"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.259119"},{"key":"e_1_2_7_4_1","unstructured":"\u00a0"},{"key":"e_1_2_7_5_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj3400711"},{"key":"e_1_2_7_6_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj20011714"},{"key":"e_1_2_7_7_1","unstructured":"\u00a0"},{"key":"e_1_2_7_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560030821"},{"key":"e_1_2_7_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(90)80821-Y"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(00)00160-X"},{"key":"e_1_2_7_11_1","unstructured":"\u00a0"},{"key":"e_1_2_7_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi500934j"},{"key":"e_1_2_7_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(97)01115-8"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi027045s"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/87610"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2016.00107"},{"key":"e_1_2_7_17_1","unstructured":"\u00a0"},{"key":"e_1_2_7_18_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7CP00277G"},{"key":"e_1_2_7_19_2","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/21.11.2509"},{"key":"e_1_2_7_20_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.035436.108"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja310165u"},{"key":"e_1_2_7_22_1","unstructured":"\u00a0"},{"key":"e_1_2_7_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2009.07.002"},{"key":"e_1_2_7_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2009.05.025"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00791a014"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi801786y"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)55681-7"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00726-015-2061-7"},{"key":"e_1_2_7_29_1","unstructured":"\u00a0"},{"key":"e_1_2_7_30_2","first-page":"255","volume":"86","author":"Vergara F.","year":"1973","journal-title":"Trans. Assoc. Am. Physicians"},{"key":"e_1_2_7_31_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.183.4120.81"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.21334"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1996.0897"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1021\/ct200133y"},{"key":"e_1_2_7_35_1","unstructured":"\u00a0"},{"key":"e_1_2_7_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(03)74017-8"},{"key":"e_1_2_7_37_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg068"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2009.07.003"},{"key":"e_1_2_7_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/0263-7855(96)00018-5"},{"key":"e_1_2_7_40_1","unstructured":"M. Frisch G. Trucks H. Schlegel G. Scuseria M. Robb J. Cheeseman G. Scalmani V. Barone B. Mennucci G. Petersson Gaussian Inc. Wallingford CT2009."},{"key":"e_1_2_7_41_1","unstructured":"\u00a0"},{"key":"e_1_2_7_42_2","doi-asserted-by":"publisher","DOI":"10.2174\/0929867311320180002"},{"key":"e_1_2_7_43_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.21082"},{"key":"e_1_2_7_44_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-319-09976-7_11"},{"key":"e_1_2_7_45_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.21256"},{"key":"e_1_2_7_46_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1996.0897"},{"key":"e_1_2_7_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(95)80037-9"},{"key":"e_1_2_7_48_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.540050204"},{"key":"e_1_2_7_49_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20035"},{"key":"e_1_2_7_50_1","unstructured":"D.\u2005A. Case T.\u2005A. Darden T.\u2005E. Cheatham C.\u2005L. Simmerling J. Wang R.\u2005E. Duke R. Luo R.\u2005C. Walker W. Zhang K.\u2005M. Merz B. Roberts S. Hayik A. Roitberg G. Seabra J. Swails A.\u2005W. Goetz I. Kolossv\u00e1ry K.\u2005F. Wong F. Paesani J. Vanicek R.\u2005M. Wolf J. Liu X. Wu S.\u2005R. Brozell T. Steinbrecher H. Gohlke Q. Cai X. Ye J. Wang M.\u2005J. Hsieh G. Cui D.\u2005R. Roe D.\u2005H. Mathews M.\u2005G. Seetin R. Salomon-Ferrer C. Sagui V. Babin T. Luchko S. Gusarov A. Kovalenko P.\u2005A. Kollman AMBER 12 University of California San Francisco 2012."},{"key":"e_1_2_7_51_1","unstructured":"\u00a0"},{"key":"e_1_2_7_52_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.22711"},{"key":"e_1_2_7_53_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.21123"},{"key":"e_1_2_7_54_1","doi-asserted-by":"publisher","DOI":"10.1063\/1.481505"},{"key":"e_1_2_7_55_1","doi-asserted-by":"publisher","DOI":"10.1063\/1.2464095"},{"key":"e_1_2_7_56_1","unstructured":"\u00a0"},{"key":"e_1_2_7_57_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.540160911"},{"key":"e_1_2_7_58_2","doi-asserted-by":"publisher","DOI":"10.1002\/cphc.201700339"},{"key":"e_1_2_7_59_2","volume":"7","author":"Sousa S. F.","year":"2016","journal-title":"Wiley Interdiscip. Rev.: Comput. Mol. Sci."},{"key":"e_1_2_7_60_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.25189"},{"key":"e_1_2_7_61_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cplett.2004.06.011"},{"key":"e_1_2_7_62_1","doi-asserted-by":"publisher","DOI":"10.1021\/ar00072a001"},{"key":"e_1_2_7_63_1","unstructured":"\u00a0"},{"key":"e_1_2_7_64_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.4821834"},{"key":"e_1_2_7_65_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.4773581"},{"key":"e_1_2_7_66_1","unstructured":"\u00a0"},{"key":"e_1_2_7_67_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jctc.5b00359"},{"key":"e_1_2_7_68_2","doi-asserted-by":"publisher","DOI":"10.1021\/ct501129s"},{"key":"e_1_2_7_69_1","unstructured":"\u00a0"},{"key":"e_1_2_7_70_2","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201900548"},{"key":"e_1_2_7_71_2","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.8b02321"},{"key":"e_1_2_7_72_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jctc.8b00348"},{"key":"e_1_2_7_73_1","first-page":"8","author":"Neese F.","year":"2017","journal-title":"Wiley Interdiscip. Rev.: Comput. Mol. Sci."},{"key":"e_1_2_7_74_1","unstructured":"\u00a0"},{"key":"e_1_2_7_75_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.456153"},{"key":"e_1_2_7_76_2","doi-asserted-by":"publisher","DOI":"10.1039\/B415208E"},{"key":"e_1_2_7_77_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.1445115"},{"key":"e_1_2_7_78_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0009-2614(98)00866-5"},{"key":"e_1_2_7_79_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.9b00739"}],"container-title":["ChemPhysChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cphc.202000751","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cphc.202000751","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cphc.202000751","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T21:35:50Z","timestamp":1760391350000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cphc.202000751"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,19]]},"references-count":79,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2021,4,19]]}},"alternative-id":["10.1002\/cphc.202000751"],"URL":"https:\/\/doi.org\/10.1002\/cphc.202000751","archive":["Portico"],"relation":{},"ISSN":["1439-4235","1439-7641"],"issn-type":[{"value":"1439-4235","type":"print"},{"value":"1439-7641","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,19]]},"assertion":[{"value":"2020-09-01","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-03-19","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}