期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1999
卷号：96
期号：4
页码：1262-1267
DOI：10.1073/pnas.96.4.1262
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Hen egg-white lysozyme dissolved in glycerol containing 1% water was studied by using CD and amide proton exchange monitored by two-dimensional 1H NMR. The far- and near-UV CD spectra of the protein showed that the secondary and tertiary structures of lysozyme in glycerol were similar to those in water. Thermal melting of lysozyme in glycerol followed by CD spectral changes indicated unfolding of the tertiary structure with a Tm of 76.0 {+/-} 0.2{degrees}C and no appreciable loss of the secondary structure up to 85{degrees}C. This is in contrast to the coincident denaturation of both tertiary and secondary structures with Tm values of 74.8 {+/-} 0.4{degrees}C and 74.3 {+/-} 0.7{degrees}C, respectively, under analogous conditions in water. Quenched amide proton exchange experiments revealed a greater structural protection of amide protons in glycerol than in water for a majority of the slowly exchanging protons. The results point to a highly ordered, native-like structure of lysozyme in glycerol, with the stability exceeding that in water.
