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Analysing Cytochrome c Aggregation and Fibrillation upon Interaction with Acetonitrile: an in Vitro Study

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Abstract

The propensity of native state to form aggregated and fibrillar assemblies is a hallmark of amyloidosis. Our study was focused at analyzing the aggregation and fibrillation tendency of cytochrome c in presence of an organic solvent i.e. acetonitrile. In vitro analysis revealed that the interaction of cytochrome c with acetonitrile facilitated the oligomerization of cytochrome c via the passage through an intermediate state which was obtained at 20 % v/v concentration of acetonitrile featured by a sharp hike in the ANS fluorescence intensity with a blue shift of 20 nm compared to the native state. Oligomers and fibrils were formed at 40 and 50 % v/v concentration respectively as indicated by a significant hike in the ThT fluorescence intensity, red shift of 55 nm in congo red binding assay and an increase in absorbance at 350 nm. They possess β-sheet structure as evident from appearance of peak at 217 nm. Finally, authenticity of oligomeric and fibrillar species was confirmed by TEM imaging which revealed bead like aggregates and a meshwork of thread like fibrils respectively. It could be suggested that the fibrillation of bovine cytchrome c could serve as a model protein to unravel the general aggregation and fibrillation pattern of heme proteins.

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Acknowledgments

The authors are highly thankful for the facilities available at AMU Aligarh. Financial support in the form of an ICMR project, ICMR [45/3/2014-BIO/BMS] is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, UP, 202 002, India

    Mohammad Furkan, Naveed Ahmad Fazili, Mohammad Afsar & Aabgeena Naeem

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  1. Mohammad Furkan
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  2. Naveed Ahmad Fazili
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  3. Mohammad Afsar
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  4. Aabgeena Naeem
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Correspondence to Aabgeena Naeem.

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Mohammad Furkan and Naveed Ahmad Fazili have equal contribution

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Furkan, M., Fazili, N.A., Afsar, M. et al. Analysing Cytochrome c Aggregation and Fibrillation upon Interaction with Acetonitrile: an in Vitro Study. J Fluoresc 26, 1959–1966 (2016). https://doi.org/10.1007/s10895-016-1889-x

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  • DOI: https://doi.org/10.1007/s10895-016-1889-x

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