Folding and Unfolding Kinetics of Unpurified Proteins by Pulse Proteolysis

Title:Folding and Unfolding Kinetics of Unpurified Proteins by Pulse Proteolysis

Volume: 23 Issue: 11

Author(s): Kanako Shima, Jun Okada, Satoshi Sano and Kazufumi Takano

Affiliation:

Keywords: Cell lysate, ribonuclease H2, subtilisin, Thermococcus kodakaraensis, circular dichroism, guanidine hydrochloride, tricine-SDS-PAGE.

Abstract: It has been reported that pulse proteolysis may be used to investigate protein unfolding kinetics in cell lysate. However, the method has not become popular and we could not judge whether or not it is effective for protein folding study. In this work, we examined the folding and unfolding kinetics of a protein and its variants without purification by pulse proteolysis. The unfolding and refolding rates of the unpurified proteins were similar to those of the purified proteins determined by pulse proteolysis and circular dichroism. Furthermore, because we used a super-stable subtilisin as a protease, we could evaluate the kinetics at 50°C. The present work demonstrates the validity of pulse proteolysis for folding and unfolding studies of unpurified proteins.

Cite this article as:

Shima Kanako, Okada Jun, Sano Satoshi and Takano Kazufumi, Folding and Unfolding Kinetics of Unpurified Proteins by Pulse Proteolysis, Protein & Peptide Letters 2016; 23 (11) . https://dx.doi.org/10.2174/0929866523666160921125257