摘要
It is known that urea and guanidine hydrochloride(GuHC1) induce conformational change of proteins in a certain range of molar ratios. In our research, a-naphthylamine(NA) above 10^-4 mol/L at pH 7.0 was discovered to perturb the conformation of CopC, a copper resistant protein with a Greek fl-barrel motif; this was reflected by the greater fluorescence quenching and red-shifted emission peak of CopC. The conformation change of CopC was also verified in acrylamide collision experiment by comparing quenching levels of CopC in the presence or the abscence of NA. Comparison of emission change of CopC-Cu^2+ with that of apo-CopC showed a consistent result with their denaturation in GuHCl. And decreasing fluorescence polarization of CopC with increasing NA concentration is consistent with a more extend conformation of CopC. Interaction mechanism was also explored.
It is known that urea and guanidine hydrochloride(GuHC1) induce conformational change of proteins in a certain range of molar ratios. In our research, a-naphthylamine(NA) above 10^-4 mol/L at pH 7.0 was discovered to perturb the conformation of CopC, a copper resistant protein with a Greek fl-barrel motif; this was reflected by the greater fluorescence quenching and red-shifted emission peak of CopC. The conformation change of CopC was also verified in acrylamide collision experiment by comparing quenching levels of CopC in the presence or the abscence of NA. Comparison of emission change of CopC-Cu^2+ with that of apo-CopC showed a consistent result with their denaturation in GuHCl. And decreasing fluorescence polarization of CopC with increasing NA concentration is consistent with a more extend conformation of CopC. Interaction mechanism was also explored.
基金
Supported by the National Natural Science Foundation of China(No. 20771068)
the Natural Science Foundation of Shanxi Province, China(No. 2007011024)