{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T00:24:46Z","timestamp":1761611086955},"reference-count":53,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2009,4,13]],"date-time":"2009-04-13T00:00:00Z","timestamp":1239580800000},"content-version":"vor","delay-in-days":2993,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2001,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Influence of isopropanol (iPrOH) on the structural dynamics of <jats:italic>Thermomyces lanuginosa<\/jats:italic> lipase (TLL) was studied by steady\u2010state, time\u2010resolved, and stopped\u2010flow fluorescence spectroscopy, monitoring the intrinsic emission of Trp residues. The fluorescence of the four Trps of the wild\u2010type enzyme report on the global changes of the whole lipase molecule. To monitor the conformational changes in the so\u2010called \u201clid,\u201d an \u03b1\u2010helical surface loop, the single Trp mutant W89m (W117F, W221H, W260H) was employed. Circular dichroism (CD) spectra revealed that iPrOH does not cause major alterations in the secondary structures of the wild\u2010type TLL and W89m. With increasing [iPrOH], judged by the ratio of emission intensities at 350 nm and 330 nm, the average microenvironment of the Trps in the wild\u2010type TLL became more hydrophobic, whereas Trp89 of W89m moved into a more hydrophilic microenvironment. Time\u2010resolved fluorescence measurements revealed no major changes to be induced by iPrOH neither in the shorter fluorescence lifetime component (\u03c4<jats:sub>1<\/jats:sub> = 0.5\u20131.2 ns) for the wild\u2010type TLL nor in the longer fluorescence lifetime component (\u03c4<jats:sub>2<\/jats:sub> = 4.8\u20136.0 ns) in the wild\u2010type TLL and the W89m mutant. Instead, for W89m on increasing iPrOH from 25% to 50% the value for \u03c4<jats:sub>1<\/jats:sub> increased significantly, from 0.43 to 1.5 ns. The shorter correlation time \u03d5<jats:sub>1<\/jats:sub> of W89m had a minimum of 0.08 ns in 25% iPrOH. Judged from the residual anisotropy r<jats:sub>\u221e<\/jats:sub> the amplitude of the local motion of Trp89 increased upon increasing [iPrOH] 10%. Stopped\u2010flow fluorescence spectroscopy measurements suggested the lid to open within \u22482 ms upon transfer of W89m into 25% iPrOH. Steady\u2010state anisotropies and longer correlation times revealed increasing concentrations of iPrOH to result also in the formation of dimers as well as possibly also higher oligomers by TLL.<\/jats:p>","DOI":"10.1110\/ps.21201","type":"journal-article","created":{"date-parts":[[2002,7,28]],"date-time":"2002-07-28T22:41:58Z","timestamp":1027896118000},"page":"339-351","source":"Crossref","is-referenced-by-count":26,"title":["Effects of i\u2010propanol on the structural dynamics of <i>Thermomyces lanuginosa<\/i> lipase revealed by tryptophan fluorescence"],"prefix":"10.1002","volume":"10","author":[{"given":"Keng","family":"Zhu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arimatti","family":"Jutila","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Esa K.J.","family":"Tuominen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paavo K.J.","family":"Kinnunen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2009,4,13]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi972998p"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/343767a0"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2744(68)90078-8"},{"key":"e_1_2_8_6_1","first-page":"4965","article-title":"Catalysis by adsorbed enzymes. 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