Abstract
This protocol aims to evaluate folding status of proteins, utilizing trypsin sensitivity. Unfolded/misfolded proteins are more susceptible to trypsin than folded proteins, because trypsin easily accesses and cleaves loosely folded parts of proteins. This method is especially useful to compare tightness of the folding among wild-type and mutant proteins. As trypsin generally cleaves a peptide bond at the carboxyl-terminal side of the amino acids lysine or arginine, this method can be used to analyze the folding status of different types of proteins such as integral membrane or soluble proteins (Ninagawa et al., 2015) and is applicable to cell lysates of any species and tissues as well as to recombinant proteins. You can use this technique with regular molecular and cell biology equipment.
Keywords: Trypsin, Protein folding, Integral membrane and soluble proteins
Materials and Reagents
Equipment
Software
Procedure
Notes
Recipes
Acknowledgments
This protocol was adapted from and used in Ninagawa et al. (2015), Izawa et al. (2012) and Xu et al. (2013).
References
Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol. You are highly recommended to post your data including images for the troubleshooting.
You are highly recommended to post your data including images for the troubleshooting.