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Incubation of the inactive enzyme at pH values \u22656.5 and 25\u00b0C resulted in a fast and partial reactivation of the enzyme, which occurred with slow onset of steady state during enzymatic turnover. The rate of approaching steady state was independent of the concentrations of choline and enzyme, but increased to a limiting value with increasing pH, defining a p<jats:italic>K<\/jats:italic><jats:sub>a<\/jats:sub> value of \u223c7.3 for an unprotonated group required for enzyme activation. Prolonged incubation of the inactive enzyme at pH 6 and temperatures \u226520\u00b0C, at which no hysteretic behavior was observed, resulted in the slow and full recovery of activity over 3 h, associated with a conformational change that reverted the enzyme to the native form. Activation of the enzyme at pH 6 was enthalpy\u2010driven with \u0394<jats:italic>H<\/jats:italic><jats:sup>\u2021<\/jats:sup> and T\u0394<jats:italic>S<\/jats:italic><jats:sup>\u2021<\/jats:sup> values of \u223c112 kJ mol<jats:sup>\u22121<\/jats:sup> and \u223c20 kJ mol<jats:sup>\u22121<\/jats:sup> determined at 25\u00b0C. These data suggest that freezing the enzyme at low pH induces a localized and reversible conformational change that is associated with the complete and reversible loss of catalytic activity. Proteins 2007. \u00a9 2006 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/prot.21238","type":"journal-article","created":{"date-parts":[[2006,12,1]],"date-time":"2006-12-01T22:47:25Z","timestamp":1165013245000},"page":"611-620","source":"Crossref","is-referenced-by-count":9,"title":["Trapping choline oxidase in a nonfunctional conformation by freezing at low pH"],"prefix":"10.1002","volume":"66","author":[{"given":"Jane V.","family":"Hoang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanni","family":"Gadda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2006,12]]},"reference":[{"key":"e_1_2_7_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)93476-9"},{"key":"e_1_2_7_3_2","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1016\/S0021-9258(18)63288-0","article-title":"Effect of freezing and thawing on the 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