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Conformational changes in mutant lysozymes detected with monoclonal antibody
Author(s) -
Kato Akio,
Shimizu Toshiaki,
Saga Shinichiro
Publication year - 1995
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 - 10.1016/0014-5793(95)00846-2
Subject(s) - lysozyme , mutant , monoclonal antibody , chemistry , denaturation (fissile materials) , conformational change , biochemistry , guanidine , microbiology and biotechnology , biophysics , antibody , biology , immunology , nuclear chemistry , gene
A monoclonal antibody (mAb) against hen egg white lysozyme (HEWL) with the exquisitely sensitive specificity to native conformation was prepared to detect the conformational changes in mutant lysozymes constructed by genetic modification in a yeast expression system. The binding of mAb with lysozyme was decreased both by denaturation with heat and guanidine‐HCl, corresponding to the denaturation curves of lysozyme. These results demonstrate that mAb is a powerful probe to monitor the conformational changes in the lysozyme molecule. By using this probe, the conformational change of various mutant lysozymes was detected. A good correlation was observed between the binding with mAb and the ΔG (Gibbs free energy change), reflecting the conformational stability of wild‐type and seven mutant lysozymes. This result suggests that a monoclonal antibody with the specificity for native conformation can be used as a powerful prove of protein conformation.