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Study of Cosolvent-Induced α-Chymotrypsin Fibrillogenesis: Does Protein Surface Hydrophobicity Trigger Early Stages of Aggregation Reaction?

The Protein Journal volume 28, Article number: 349 (2009) Cite this article

Abstract

The misfolding of specific proteins is often associated with their assembly into fibrillar aggregates, commonly termed amyloid fibrils. Despite the many efforts expended to characterize amyloid formation in vitro, there is no deep knowledge about the environment (in which aggregation occurs) as well as mechanism of this type of protein aggregation. Alpha-chymotrypsin was recently driven toward amyloid aggregation by the addition of intermediate concentrations of trifluoroethanol. In the present study, approaches such as turbidimetric, thermodynamic, intrinsic fluorescence and quenching studies as well as chemical modification have been successfully used to elucidate the underlying role of hydrophobic interactions (involved in early stages of amyloid formation) in α-chymotrypsin-based experimental system.

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Abbreviations

BSA:

Bovine serum albumin

DSC:

Differential Scanning Calorimetry

PSH:

Protein surface hydrophobicity

TFE:

2,2,2-Trifluoroethanol

CR:

Congo red

ThT:

Thioflavin T

α-CD:

α-Cyclodextrin

TNBS:

2,4,6-Trinitrobenzenesulfonic acid

ANS:

1,8-Anilinonaphtalenesulfonate

BTEE:

N-Benzoyl-l-tyrosyl-ethyl ester

PMSF:

Phenyl methyl sulphonyl fluoride

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Acknowledgments

We thank Mr. M. Beyrami, Mr. F. Naderi and Dr. H. Adibi for their invaluable helps. Also, effective assistance provided by Ms M. Mehrabi and gifts (BTEE and α-chymotrypsin) from Dr. R. H. Sajedi, are gratefully acknowledged. This publication was made possible by grants from the “Pharmaceutical Sciences Research Network” of I. R. Iran and Kermanshah University of Medical Sciences. The sponsors, however, had any direct involvement with this work. The authors are also deeply indebted to Dr. A. A. Moosavi-Movahedi (Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran) for his help in performing DSC measurements. This work was supported in part by a grant to R. K. from the “Pharmaceutical Sciences Research Network” of I. R. Iran.

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Affiliations

  1. Department of Pharmacognosy and Biotechnology, Faculty of Pharmacy, Kermanshah University of Medical Sciences, P. O. Box 67145-1673, Kermanshah, Iran

    Reza Khodarahmi & Hosnieh Soori

  2. Medical Biology Research Center, Kermanshah University of Medical Sciences, P. O. Box 67155-1616, Kermanshah, Iran

    Reza Khodarahmi

  3. Faculty of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Mojtaba Amani

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  1. Reza Khodarahmi
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  2. Hosnieh Soori
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  3. Mojtaba Amani
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Correspondence to Reza Khodarahmi.

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Khodarahmi, R., Soori, H. & Amani, M. Study of Cosolvent-Induced α-Chymotrypsin Fibrillogenesis: Does Protein Surface Hydrophobicity Trigger Early Stages of Aggregation Reaction?. Protein J 28, 349 (2009). https://doi.org/10.1007/s10930-009-9200-5

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  • DOI: https://doi.org/10.1007/s10930-009-9200-5

Keywords

  • Fluorescence
  • α-Chymotrypsin
  • Aggregation
  • TFE
  • Hydrophobicity