{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T18:34:48Z","timestamp":1760985288117},"reference-count":46,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2015,1,30]],"date-time":"2015-01-30T00:00:00Z","timestamp":1422576000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Science Council of Lithuania","award":["VP1- 3.1.-SMM-07-K-02-009"],"award-info":[{"award-number":["VP1- 3.1.-SMM-07-K-02-009"]}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2015,3]]},"abstract":"<jats:p>Carbonic anhydrase (<jats:styled-content style=\"fixed-case\">CA<\/jats:styled-content>) <jats:styled-content style=\"fixed-case\">VI<\/jats:styled-content> is a potential drug target for cariogenesis and cancer of the salivary gland. It is the only secreted human <jats:styled-content style=\"fixed-case\">CA<\/jats:styled-content> isozyme which is found in saliva and milk. Here, <jats:styled-content style=\"fixed-case\">CA VI<\/jats:styled-content> was expressed in bacterial and mammalian cell cultures and directly affinity\u2010purified from human saliva. The binding of 4\u2010substituted\u20102,3,5,6\u2010tetrafluorobenezenesulfonamides to the native and recombinant <jats:styled-content style=\"fixed-case\">CA VI<\/jats:styled-content> from these three sources was compared. Interaction between the enzyme and inhibitors was determined by fluorescent thermal shift assay and isothermal titration calorimetry. The observed dissociation constants were the same within the error margin for all three <jats:styled-content style=\"fixed-case\">CA VI<\/jats:styled-content> preparations. The intrinsic binding parameters for the compounds were obtained by determining and dissecting the binding\u2010linked protonation reactions. Intrinsic thermodynamic parameters of binding arrange the compounds in a buffer\u2010 and <jats:styled-content style=\"fixed-case\">pH<\/jats:styled-content>\u2010independent manner. Intrinsic binding constants of nonfluorinated compounds were significantly stronger than those of fluorinated benzenesulfonamides. An opposite result was determined for the observed binding constants. The increase in observed affinity of the fluorinated compounds was due to the fluorine effect on diminishing the <jats:styled-content style=\"fixed-case\">p<jats:italic>K<\/jats:italic><\/jats:styled-content><jats:sub>a<\/jats:sub> of the compounds but not due to direct recognition of the protein. The temperature\u2013stability profiles for recombinant and native <jats:styled-content style=\"fixed-case\">CA VI<\/jats:styled-content> were compared and showed that <jats:styled-content style=\"fixed-case\">CA VI<\/jats:styled-content> is more stable in slightly acidic than neutral conditions.<\/jats:p>","DOI":"10.1111\/febs.13196","type":"journal-article","created":{"date-parts":[[2015,1,14]],"date-time":"2015-01-14T19:07:44Z","timestamp":1421262464000},"page":"972-983","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Intrinsic binding of 4\u2010substituted\u20102,3,5,6\u2010tetrafluorobenezenesulfonamides to native and recombinant human carbonic anhydrase VI"],"prefix":"10.1111","volume":"282","author":[{"given":"Justina","family":"Kazokait\u0117","sequence":"first","affiliation":[{"name":"Department of Biothermodynamics and Drug Design Institute of Biotechnology Vilnius University Lithuania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Goda","family":"Milinavi\u010di\u016bt\u0117","sequence":"additional","affiliation":[{"name":"Department of Biothermodynamics and Drug Design Institute of Biotechnology Vilnius University Lithuania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joana","family":"Smirnovien\u0117","sequence":"additional","affiliation":[{"name":"Department of Biothermodynamics and Drug Design Institute of Biotechnology Vilnius University Lithuania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jurgita","family":"Matulien\u0117","sequence":"additional","affiliation":[{"name":"Department of Biothermodynamics and Drug Design Institute of Biotechnology Vilnius University Lithuania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daumantas","family":"Matulis","sequence":"additional","affiliation":[{"name":"Department of Biothermodynamics and Drug Design Institute of Biotechnology Vilnius University Lithuania"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,1,30]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrd2467"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2012.04.044"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(79)80634-1"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)91031-8"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1159\/000016762"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)75797-4"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1177\/38.7.2113069"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1177\/41.3.8429198"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.121172598"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1177\/002215540205000608"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1369\/jhc.4A6282.2004"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1292\/jvms.68.131"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0722.2006.00273.x"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm0612057"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1159\/000016514"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.14219\/jada.archive.2003.0367"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1018889120034"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1998.9356"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.3945\/ajcn.2010.29418"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12929-014-0082-2"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja01167a012"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/cr200176r"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1747-0285.2009.00902.x"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2012.02.038"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-8-17"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmr.2330"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi048135v"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms10062662"},{"key":"e_1_2_8_30_1","first-page":"247","volume-title":"Protein Thermal Denaturation Measurements Via a Fluorescent Dye. 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