Unfolding During Urea Denaturation of a Low Molecular Weight Phytocystatin (Thiol Protease Inhibitor) Purified from Phaseolus mungo (Urd)

Title: Unfolding During Urea Denaturation of a Low Molecular Weight Phytocystatin (Thiol Protease Inhibitor) Purified from Phaseolus mungo (Urd)

Volume: 13 Issue: 4

Author(s): Sandeep Sharma, Fouzia Rashid and Bilqees Bano

Affiliation:

Keywords: Plant cystatins, Fluorescence Spectroscopy, Purification, Proteins, Thiol protease inhibitors, Circular Dichroism

Abstract: In the present study, two phytocystatins were purified to homogeneity as peaks I and II with molecular weights of 19 kDa and 17 kDa, respectively, as determined by SDS-PAGE and mass spectrometry. Both PMCs I and II were purified with a greater than 1000-fold purification and overall yield of about 16-18%. The effect of urea on PMC I and II was analysed by fluorescence and Circular Dichroism (CD) spectroscopy. Fluorescence studies suggest a red shift of the maximum emission at higher urea concentrations. PMC I and II are extremely stable protein inhibitors with regards to temperature and pH stability. FTIR studies show predominant a-helical structure in both the cystatins. CD analysis results show change in urea concentration-dependent loss in ellipticity, as well as in the shape of the CD spectrum compared to the intact phytocystatin.

Cite this article as:

Sharma Sandeep, Rashid Fouzia and Bano Bilqees, Unfolding During Urea Denaturation of a Low Molecular Weight Phytocystatin (Thiol Protease Inhibitor) Purified from Phaseolus mungo (Urd), Protein & Peptide Letters 2006; 13 (4) . https://dx.doi.org/10.2174/092986606775974447

DOI https://dx.doi.org/10.2174/092986606775974447	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305