{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,20]],"date-time":"2025-09-20T20:23:53Z","timestamp":1758399833602},"reference-count":29,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2001,12,25]],"date-time":"2001-12-25T00:00:00Z","timestamp":1009238400000},"content-version":"vor","delay-in-days":693,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[2000,2]]},"abstract":"<jats:p>We analysed the conformational states of free, <jats:italic>tet<\/jats:italic> operator\u2010bound and anhydrotetracycline\u2010bound Tet repressor employing a Trp\u2010scanning approach. The two wild\u2010type Trp residues in Tet repressor were replaced by Tyr or Phe and single Trp residues were introduced at each of the positions 162\u2013173, representing part of an unstructured loop and the N\u2010terminal six residues of \u03b1\u2010helix 9. All mutants retained <jats:italic>in vivo<\/jats:italic> inducibility, but anhydrotetracycline\u2010binding constants were decreased up to 7.5\u2010fold when Trp was in positions 169, 170 and 173. Helical positions (168\u2013173) differed from those in the loop (162\u2013167) in terms of their fluorescence emission maxima, quenching rate constants with acrylamide and anisotropies in the free and <jats:italic>tet<\/jats:italic> operator\u2010complexed proteins. Trp fluorescence emission decreased drastically upon atc binding, mainly due to energy transfer. For all proteins, either free, <jats:italic>tet<\/jats:italic> operator bound or anhydrtetracycline\u2010bound, mean fluorescence lifetimes were determined to derive quenching rate constants. Solvent\u2010accessible surfaces of the respective Trp side chains were calculated and compared with the quenching rate constants in the anhydrotetracycline\u2010bound complexes. The results support a model, in which residues in the loop become more exposed, whereas residues in \u03b1\u2010helix 9 become more buried upon the induction of TetR by anhydrotetracycline.<\/jats:p>","DOI":"10.1046\/j.1432-1327.2000.01063.x","type":"journal-article","created":{"date-parts":[[2003,3,11]],"date-time":"2003-03-11T18:00:40Z","timestamp":1047405640000},"page":"821-829","source":"Crossref","is-referenced-by-count":19,"title":["Trp scanning analysis of Tet repressor reveals conformational changes associated with operator and anhydrotetracycline binding"],"prefix":"10.1111","volume":"267","author":[{"given":"Martin","family":"Kintrup","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Schubert","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Kunz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie","family":"Chabbert","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia","family":"Alberti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa","family":"Bombarda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siegfried","family":"Schneider","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wolfgang","family":"Hillen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2001,12,25]]},"reference":[{"key":"e_1_2_7_2_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.mi.48.100194.002021"},{"key":"e_1_2_7_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(91)90089-C"},{"key":"e_1_2_7_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7792603"},{"key":"e_1_2_7_5_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.14.2723"},{"key":"e_1_2_7_6_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.8153629"},{"key":"e_1_2_7_7_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1994.0138"},{"key":"e_1_2_7_8_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1998.1775"},{"key":"e_1_2_7_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsb0895-693"},{"key":"e_1_2_7_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1991.tb04992.x"},{"key":"e_1_2_7_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1988.tb03290.x"},{"key":"e_1_2_7_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.175.4.1206-1210.1993"},{"key":"e_1_2_7_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/0021-9673(96)00232-4"},{"key":"e_1_2_7_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(90)90351-Q"},{"key":"e_1_2_7_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(92)91065-W"},{"key":"e_1_2_7_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi952683e"},{"key":"e_1_2_7_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(88)90120-9"},{"key":"e_1_2_7_18_2","volume-title":"Experiments in Molecular Genetics","author":"Miller J.H.","year":"1972"},{"key":"e_1_2_7_19_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4615-7658-7"},{"key":"e_1_2_7_20_2","volume-title":"Topics in Fluorescence Spectroscopy. 1 Techniques.","author":"Lakowicz J.R.","year":"1991"},{"key":"e_1_2_7_21_2","unstructured":"Sali A.(1995) Modeller.Rockefeller University New York."},{"key":"e_1_2_7_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.340110407"},{"key":"e_1_2_7_23_2","doi-asserted-by":"crossref","first-page":"12269","DOI":"10.1016\/S0021-9258(18)45346-X","article-title":"Tryptophan in alpha\u2010helix 3 of Tet repressor forms a sequence\u2010specific contact with tet operator in solution.","volume":"262","author":"Hansen D.","year":"1987","journal-title":"J. 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