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Oxidative refolding of lysozyme in trifluoroethanol (TFE) and ethylene glycol: interfering role of preexisting α‐helical structure and intermolecular hydrophobic interactions
Author(s) -
Prabha C.Ratna,
Mohan Rao Ch.
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)01442-x
Subject(s) - lysozyme , chemistry , ethylene glycol , circular dichroism , folding (dsp implementation) , protein folding , hydrophobic effect , protein secondary structure , oxidative folding , oxidative phosphorylation , intermolecular force , photochemistry , crystallography , biophysics , organic chemistry , disulfide bond , biochemistry , molecule , protein disulfide isomerase , electrical engineering , biology , engineering
The oxidative refolding of equilibrium intermediates of lysozyme stabilized in trifluoroethanol (TFE) and ethylene glycol was monitored. Equilibrium intermediates of disulfide reduced lysozyme in TFE are known to contain considerable amounts of α‐helical structure and resemble the early intermediate in the oxidative refolding of lysozyme. We find that the intermediates in TFE do not proceed to folding; they form aggregates. However, interestingly, intermediates in ethylene glycol refold to the native state with improved folding yield. Secondary structure of these intermediates was monitored by far‐UV circular dichroism. Our results indicate that formation of α‐helical structure prior to oxidative refolding does not help the process in the case of lysozyme. Interfering with intermolecular hydrophobic interactions in the unfolded state is more productive.