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Addition of methanol induced no global change in the native conformation itself, but induced a transition from the native state to the denatured state which was highly cooperative, as shown by the coincidence of transition curves monitored by the far\u2010and near\u2010UV CD spectroscopy, by isodichroic points in the far\u2010 and near\u2010UV CD spectra and by the concomitant disappearance of individual <jats:sup>1<\/jats:sup>H NMR signals of the native state. The ANS binding experiments could detect no intermediate conformer similar to the molten globule state in the process of the methanol denaturation. However, at high concentration of methanol, e.g., 60% (v\/v) methanol\/water, a highly helical state (H) was realized. The H state had a helical content much higher than the native state, monitored by far\u2010UV CD spectroscopy, and had no specific tertiary structure, monitored both by near\u2010UV CD and NMR spectroscopy. The radius of gyration in the H state, 24.9 \u00c5, was significantly larger than that in the native state (15.7 \u00c5). The Kratky plot for the H state did not show a clear peak and was quite similar to that for the urea\u2010denatured state, indicating a complete lack of globularity. Thus we conclude that the H state has a considerably expanded, flexible broken rod\u2010like conformation which is clearly distinguishable from the \u201cmolten globule\u201d state. The stability of both N and H states depends on p<jats:sup>H<\/jats:sup> and methanol concentration. Thus a phase diagram involving N and H was constructed.<\/jats:p>","DOI":"10.1002\/pro.5560070317","type":"journal-article","created":{"date-parts":[[2010,7,12]],"date-time":"2010-07-12T05:31:53Z","timestamp":1278912713000},"page":"681-688","source":"Crossref","is-referenced-by-count":55,"title":["The methanol\u2010induced transition and the expanded helical conformation in hen lysozyme"],"prefix":"10.1002","volume":"7","author":[{"given":"Yuji 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