Molecular Crowding Affects the Conformational Fluctuations, Peroxidase Activity, and Folding Landscape of Yeast Cytochrome c
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Molecular Crowding Affects the Conformational Fluctuations,
Peroxidase Activity, and Folding Landscape of Yeast Cytochrome c
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| Creator |
Paul, Simanta Sarani
Sil, Pallabi Chakraborty, Ritobrita Haldar, Shubhasis Chattopadhyay, Krishnananda |
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| Subject |
Structural Biology & Bioinformatics
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| Description |
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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| Publisher |
American Chemical Society
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| Date |
2016
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/2536/1/BIOCHEMISTRY_.pdf
Paul, Simanta Sarani and Sil, Pallabi and Chakraborty, Ritobrita and Haldar, Shubhasis and Chattopadhyay, Krishnananda (2016) Molecular Crowding Affects the Conformational Fluctuations, Peroxidase Activity, and Folding Landscape of Yeast Cytochrome c. Biochemistry, 55 (16). pp. 2332-2343. |
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| Relation |
http://dx.doi.org/10.1021/acs.biochem.6b00053
http://www.eprints.iicb.res.in/2536/ |
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