Title
Differential domain structure stability of the severe acute respiratory syndrome coronavirus papain-like protease
Author
Ya-Wen Chou, Shu-Chun Cheng, Hsing-Yi Lai, Chi-Yuan Chou
Year
2012
Journal
Archives of Biochemistry and Biophysics
Abstract
Papain-like protease (PLpro) from severe acute respiratory syndrome (SARS) coronavirus is one of the two proteases involved in the proteolytic processing of the virion polyproteins. In addition, PLpro shows significant in vitro deubiquitinating and de-ISGylating activities. All these findings demonstrated the multifunctional nature of the PLpro. Here we report the sensitivity of PLpro to denaturant urea. An increase in urea concentration induced a reversible biphasic unfolding of the enzyme. Differently, the unfolding of the catalytic triad region located within the palm and thumb domains followed a monophasic unfolding curve. Further observations suggest that the zinc-binding domain may start to unfold during the first transition. An 80% lost of its enzymatic activity at a urea concentration lower than 1 M showed a close correlation with unfolding of the zinc-binding domain. The enzyme was also characterized in terms of hydrophobicity and size-and-shape distribution. We have demonstrated that PLpro displayed differential domain structure stability and molten globule state in its folding. These studies will not only assist in our understanding of the folding of this viral enzyme, but also that of other deubiquitinating enzymes with a similar scaffold.
Instrument
V-550, J-810
Keywords
Circular dichroism, Protein folding, Secondary structure, Chemical stability, Protein denaturation, Thermodynamics, Absorption, Kinetics, Quantitation, Biochemistry