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  Molecular Crowding Affects the Conformational Fluctuations, Peroxidase Activity, and Folding Landscape of Yeast Cytochrome c

Paul, S. S., Sil, P., Chakraborty, R., Haldar, S., & Chattopadhyay, K. (2016). Molecular Crowding Affects the Conformational Fluctuations, Peroxidase Activity, and Folding Landscape of Yeast Cytochrome c. BIOCHEMISTRY, 55(16), 2332-2343. doi:10.1021/acs.biochem.6b00053.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-E35C-6 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-E35D-4
Genre: Journal Article

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 Creators:
Paul, Simanta Sarani1, Author
Sil, Pallabi1, Author
Chakraborty, Ritobrita1, Author
Haldar, Shubhasis2, Author              
Chattopadhyay, Krishnananda1, Author
Affiliations:
1external, ou_persistent22              
2Hartl, Franz-Ulrich / Cellular Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565152              

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Free keywords: FLUORESCENCE CORRELATION SPECTROSCOPY; PROTEIN SELF-ASSOCIATION; ACID-BINDING PROTEIN; ESCHERICHIA-COLI; RELAXATION DISPERSION; CARBONIC-ANHYDRASE; AGGREGATION; STABILITY; CONFINEMENT; KINETICS
 Abstract: To understand how a protein folds and behaves inside living cells, the effects of synthetic crowding media on protein folding, function, stability, and association have been studied in detail. Because the effect of excluded volume is more prominent in an extended state than in the native protein, a majority of these studies have been conducted in the unfolded state of different model proteins. Here, we have used fluorescence correlation spectroscopy (FCS) and other biophysical methods to investigate the effect of crowding agents Ficoll70 and Dextran70 on the nativelike state of cytochrome c from yeast. Yeast cytochrome c (y-cytc) contains a substantial expanded state in its native folded condition, which is present in equilibrium with a compact conformer in aqueous buffer. We have found that the crowding medium affects the native state equilibrium between compact and expanded states, shifting its population toward the compact conformer. As a result, the peroxidase activity of y-cytc decreases. Urea-induced protein stability measurements show that the compaction destabilizes the protein due to charge repulsions between similar charged clusters. Interestingly, the time constant of conformational fluctuations between the compact and expanded conformers has been found to increase in the crowded milieu, suggesting a crucial role of the solution microviscosity.

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Language(s): eng - English
 Dates: 2016
 Publication Status: Published in print
 Pages: 12
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Degree: -

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Title: BIOCHEMISTRY
Source Genre: Journal
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Publ. Info: 1155 16TH ST, NW, WASHINGTON, DC 20036 USA : AMER CHEMICAL SOC
Pages: - Volume / Issue: 55 (16) Sequence Number: - Start / End Page: 2332 - 2343 Identifier: ISSN: 0006-2960