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Identification of folding intermediates of streblin, the most stable serine protease: biophysical analysis.
Appl Biochem Biotechnol. 2014 Jan; 172(2):658-71.AB

Abstract

Streblin, a serine proteinase from plant Streblus asper, has been used to investigate the conformational changes induced by pH, temperature, and chaotropes. The near/far UV circular dichroism activities under fluorescence emission spectroscopy and 8-aniline-1-naphthalene sulfonate (ANS) binding have been carried out to understand the unfolding of the protein in the presence of denaturants. Spectroscopic studies reveal that streblin belongs to the α+β class of proteins and exhibits stability towards chemical denaturants, guanidine hydrochloride (GuHCl). The pH-induced transition of this protein is noncooperative for transition phases between pH 0.5 and 2.5 (midpoint, 1.5) and pH 2.5 and 10.0 (midpoint, 6.5). At pH 1.0 or lower, the protein unfolds to form acid-unfolded state, and for pH 7.5 and above, protein turns into an alkaline denatured state characterized by the absence of ANS binding. At pH 2.0 (1 M GuHCl), streblin exists in a partially unfolded state with characteristics of a molten globule state. The protein is found to exhibit strong and predominant ANS binding. In total, six different intermediate states has been identified to show protein folding pathways.

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Authors+Show Affiliations

Kumar R
Molecular Biology Unit, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India, reeteshmbu@gmail.com.
Tripathi P
No affiliation info available
de Moraes FR
No affiliation info available
Caruso IP
No affiliation info available
Jagannadham MV
No affiliation info available

MeSH

Biophysical PhenomenaCircular DichroismEnzyme StabilityGuanidineHydrogen-Ion ConcentrationProtein DenaturationProtein FoldingSerine ProteasesSpectrometry, FluorescenceSpectrophotometry, UltravioletThermodynamics

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24108566

Citation

Kumar, Reetesh, et al. "Identification of Folding Intermediates of Streblin, the Most Stable Serine Protease: Biophysical Analysis." Applied Biochemistry and Biotechnology, vol. 172, no. 2, 2014, pp. 658-71.
Kumar R, Tripathi P, de Moraes FR, et al. Identification of folding intermediates of streblin, the most stable serine protease: biophysical analysis. Appl Biochem Biotechnol. 2014;172(2):658-71.
Kumar, R., Tripathi, P., de Moraes, F. R., Caruso, I. P., & Jagannadham, M. V. (2014). Identification of folding intermediates of streblin, the most stable serine protease: biophysical analysis. Applied Biochemistry and Biotechnology, 172(2), 658-71. https://doi.org/10.1007/s12010-013-0565-8
Kumar R, et al. Identification of Folding Intermediates of Streblin, the Most Stable Serine Protease: Biophysical Analysis. Appl Biochem Biotechnol. 2014;172(2):658-71. PubMed PMID: 24108566.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of folding intermediates of streblin, the most stable serine protease: biophysical analysis. AU - Kumar,Reetesh, AU - Tripathi,Pinki, AU - de Moraes,Fabio Rogerio, AU - Caruso,Icaro P, AU - Jagannadham,Medicherla V, Y1 - 2013/10/10/ PY - 2013/08/07/received PY - 2013/09/27/accepted PY - 2013/10/11/entrez PY - 2013/10/11/pubmed PY - 2014/10/8/medline SP - 658 EP - 71 JF - Applied biochemistry and biotechnology JO - Appl Biochem Biotechnol VL - 172 IS - 2 N2 - Streblin, a serine proteinase from plant Streblus asper, has been used to investigate the conformational changes induced by pH, temperature, and chaotropes. The near/far UV circular dichroism activities under fluorescence emission spectroscopy and 8-aniline-1-naphthalene sulfonate (ANS) binding have been carried out to understand the unfolding of the protein in the presence of denaturants. Spectroscopic studies reveal that streblin belongs to the α+β class of proteins and exhibits stability towards chemical denaturants, guanidine hydrochloride (GuHCl). The pH-induced transition of this protein is noncooperative for transition phases between pH 0.5 and 2.5 (midpoint, 1.5) and pH 2.5 and 10.0 (midpoint, 6.5). At pH 1.0 or lower, the protein unfolds to form acid-unfolded state, and for pH 7.5 and above, protein turns into an alkaline denatured state characterized by the absence of ANS binding. At pH 2.0 (1 M GuHCl), streblin exists in a partially unfolded state with characteristics of a molten globule state. The protein is found to exhibit strong and predominant ANS binding. In total, six different intermediate states has been identified to show protein folding pathways. SN - 1559-0291 UR - https://www.unboundmedicine.com/medline/citation/24108566/Identification_of_folding_intermediates_of_streblin_the_most_stable_serine_protease:_biophysical_analysis_ L2 - https://dx.doi.org/10.1007/s12010-013-0565-8 DB - PRIME DP - Unbound Medicine ER -