Advertisement

The Protein Journal

, Volume 36, Issue 2, pp 138–146 | Cite as

Trehalose and Magnesium Chloride Exert a Common Anti-amyloidogenic Effect Towards Hen Egg White Lysozyme

  • Rupsa Chatterjee
  • Vidyalatha Kolli
  • Nandini Sarkar
Article

Abstract

Many degenerative disorder such as Parkinsons, Alzheimers, Huntingtons disease, etc are caused due to the deposition of amyloid fibrils, formed due to the ordered aggregation of misfolded/unfolded proteins. Misfolded or unfolded proteins aggregate mostly through hydrophobic interactions which are unexposed in native state, but become exposed upon unfolding. To counteract amyloid related diseases, inhibition of the protein self assembly into fibril is a potential therapeutic strategy. The study aims at investigating the effect of selected compounds, namely trehalose and magnesium chloride hexahydrate towards inhibition and disaggregation of amyloid fibrils using Hen Egg White Lysozyme as a model. We further attempted to understand the mechanism of action with the help of various biophysical, microscopic as well as computational studies. A common mechanism of action was identified where the selected compounds exert their anti-amyloidogenic effects by altering HEWL conformations characterized by reduction in the beta sheet content and decrease in exposed hydrophobic surfaces. The altered conformation seems to have lesser amyloidogenic propensity leading to inhibition as well as disaggregation of amyloids.

Keywords

Amyloidogenesis Hen egg white lysozyme Hydrophobic Circular dichroism Thioflavin T 

Abbreviations

ANS

1-Anilinonaphthalene-8-sulphonic acid

BSA

Bovine serum albumin

CD

Circular dichroism

DDI

Distilled deionised

HEWL

Hen egg white lysozyme

PDB

Protein data bank

PTA

Phosphotungstic acid

TEM

Transmission electron microscope

ThT

Thioflavin T

Notes

Acknowledgements

The authors acknowledge the infrastructural facilities at NIT Rourkela. The fellowship provided to VK from DBT is also acknowledged.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Soto C (2001) Protein misfolding and disease; protein refolding and therapy. FEBS Lett 498:204–207CrossRefGoogle Scholar
  2. 2.
    Frare E, de Laureto PP, Zurdo J, Dobson CM, Fontana A (2004) A highly amyloidogenic region of hen lysozyme. J Mol Biol 340:1153–1165CrossRefGoogle Scholar
  3. 3.
    Dobson CM (2003) Protein folding and misfolding. Nature 426:884–890CrossRefGoogle Scholar
  4. 4.
    Sgarbossa A (2012) Natural biomolecules and protein aggregation: emerging strategies against amyloidogenesis. Int J Mol Sci 13:17121–17137CrossRefGoogle Scholar
  5. 5.
    Kelly JW (1998) The alternative conformations of amyloidogenic proteins and their multi-step assembly pathways. Curr Opin Struct Boil 8:101–106CrossRefGoogle Scholar
  6. 6.
    Avdeev MV, Aksenov VL, Gazova Z, Almasy L, Petrenko VI, Gojzewski H, Feokstystov AV, Siposova K, Antosova A, Timko M, Kopcansky P (2013) On the determination of the helical structure parameters of amyloid protofilaments by small-angle neutron scattering and atomic force microscopy. J Appl Cryst 46:224–233CrossRefGoogle Scholar
  7. 7.
    Petrenko VL, Avdeev MV, Garamus VM, Kubovcikova M, Gazova Z, Siposova K, Bulavin LA, Almasy L, Aksenov VL, Kopcansky P (2014) Structure of amyloid aggregates of lysozyme from small-angle X-ray scattering data. Phys Solid State 56:129–133CrossRefGoogle Scholar
  8. 8.
    Goldsbury C, Baxa U, Simon MN, Steven AC, Engel A, Wall JS, Aebi U, Muller SA (2011) Amyloid structure and assembly: insights from scanning transmission electron microscopy. J Struct Biol 173:1–13CrossRefGoogle Scholar
  9. 9.
    Kisilevsky R (2000) Review: amyloidogenesis—unquestioned answers and unanswered questions. J Struct Biol 130:99–108CrossRefGoogle Scholar
  10. 10.
    Sipe JD, Cohen AS (2000) Review: history of the amyloid fibril. J Struct Biol 130:88–98CrossRefGoogle Scholar
  11. 11.
    Wolfe KJ, Cyr DM (2011) Amyloid in neurodegenerative diseases: friend or foe?. Semin Cell Dev Biol 22:476–481CrossRefGoogle Scholar
  12. 12.
    Shiraki K, Kudou M, Aso Y, Takagi M (2003) Dissolution of protein aggregation by small amine compounds. Sci Technol Adv Mater 4:55–59CrossRefGoogle Scholar
  13. 13.
    Arakawa T, Ejima D, Tsumoto K, Obeyama N, Tanaka Y, Kita Y, Timasheff SN (2007) Suppression of protein interactions by arginine: a proposed mechanism of the arginine effects. Biophys Chem 127:1–8CrossRefGoogle Scholar
  14. 14.
    Ishibashi M, Tsumoto K, Tokunaga M, Ejima D, Kita Y, Arakawa T (2005) Is arginine a protein-denaturant?. Protein Expres Purify 42:1–6CrossRefGoogle Scholar
  15. 15.
    Das U, Hariprasad G, Ethayathulla AS, Manral P, Das TK, Pasha S, Mann A, Ganguli M, Verma AK, Bhat R, Chandrayan SK, Ahmed S, Sharma S, Kaur P, Singh TP, Srinivasan A (2007) Inhibition of protein aggregation: supramolecular assemblies of arginine hold the key. PLoS ONE 2:e1176CrossRefGoogle Scholar
  16. 16.
    Ignatova Z, Gierasch LM (2006) Inhibition of protein aggregation in vitro and in vivo by a natural osmoprotectant. Proc Natl Acad Sci USA 103:13357–13361CrossRefGoogle Scholar
  17. 17.
    Hamada H, Arakawa T, Shiraki K (2009) Effect of additives on protein aggregation. Curr Pharm Biotechnol 10:400–407CrossRefGoogle Scholar
  18. 18.
    Chi EY, Krishnan S, Randolph TW, Carpenter JF (2003) Physical stability of proteins in aqueous solution: mechanism and driving forces in nonnative protein aggregation. Pharm Res 20:1325–1336CrossRefGoogle Scholar
  19. 19.
    Vieira MN, Figueroa-Villar JD, Meirelles MNL, Ferreira ST, De Felice FG (2006) Small molecule inhibitors of lysozyme amyloid aggregation. Cell Biochem Biophys 44:549–553CrossRefGoogle Scholar
  20. 20.
    He J, Xing YF, Huang B, Zhang YZ, Zeng CM (2009) Tea catechins induce the conversion of preformed lysozyme amyloid fibrils to amorphous aggregates. J Agr Food Chem 57:11391–11396CrossRefGoogle Scholar
  21. 21.
    Wang SSS, Liu KN, Lee WH (2009) Effect of curcumin on the amyloid fibrillogenesis of hen egg-white lysozyme. Biophys Chem 144:78–87CrossRefGoogle Scholar
  22. 22.
    Gazova Z, Siposova K, Kurin E, Mucaji P, Nagy M (2013) Amyloid aggregation of lysozyme: the synergy study of red wine polyphenols. Proteins 81:994–1004CrossRefGoogle Scholar
  23. 23.
    He J, Wang Y, Chang AK, Xu L, Wang N, Chong X, Li H, Zhang B, Jones GW, Song Y (2014) Myricetin prevents fibrillogenesis of hen egg white lysozyme. J Agr Food Chem 62:9442–9449CrossRefGoogle Scholar
  24. 24.
    Borana MS, Mishra P, Pissurlenkar RRS, Hosur RV, Ahmad B (2014) Curcumin and kaempferol prevent lysozyme fibril formation by modulating aggregation kinetic parameters. BBA-Proteins Proteom 1844:670–680CrossRefGoogle Scholar
  25. 25.
    Singha Roy A, Utreja J, Badhei S (2015) Characterization of the binding of fisetin and morin with chicken egg lysozyme using spectroscopic and molecular docking methods. J Incl Phenom Macrocycl Chem 81:385–394CrossRefGoogle Scholar
  26. 26.
    Liu KN, Lai CM, Lee YT, Wang SN, Chen RPY, Jan JS, Liu HS, Wang SSS (2012) Curcumin’s pre-incubation temperature affects its inhibitory potency toward amyloid fibrillation and fibril-induced cytotoxicity of lysozyme. BBA-GEN Subj 1820:1774–1786CrossRefGoogle Scholar
  27. 27.
    Ueda T, Nagata M, Imoto T (2001) Aggregation and chemical reaction in hen lysozyme caused by heating at pH 6 are depressed by osmolytes, sucrose and trehalose. J Biochem 130:491–496CrossRefGoogle Scholar
  28. 28.
    Khurana R, Coleman C, Ionescu-Zanetti C, Carter SA, Krishna V, Grover RK, Roy R, Singh S (2005) Mechanism of thioflavin T binding to amyloid fibrils. J Struct Biol 151:229–238CrossRefGoogle Scholar
  29. 29.
    Ghosh S, Pandey NK, Dasgupta S (2013) (−)-Epicatechin gallate prevents alkali-salt mediated fibrillogenesis of hen egg white lysozyme. Int J Biol Macromol 54:90–98CrossRefGoogle Scholar
  30. 30.
    Lee C (2010) 1-Anilinonaphthalene-8-sulfonate (ans); a versatile fluorescent probe from protein folding study to drug design. BioWave 12:1–12Google Scholar
  31. 31.
    McCubbin WD, Kay CM, Narindrasorasak S, Kisilevsky R (1988) Circular-dichroism studies on two murine serum amyloid A proteins. Biochem J 256:775–783CrossRefGoogle Scholar
  32. 32.
    Serpell LC (2000) Alzheimer’s amyloid fibrils: structure and assembly. BBA-Mol Basis Dis 1502:16–30CrossRefGoogle Scholar
  33. 33.
    Garbuzynskiy SO, Lobanov MY, Galzitskaya OV (2010) FoldAmyloid: a method of prediction of amyloidogenic regions from protein sequence. Bioinformatics 26:326–332CrossRefGoogle Scholar
  34. 34.
    Norris R, Casey F, FitzGerald RJ, Shields D, Mooney C (2012) Predictive modelling of angiotensin converting enzyme inhibitory dipeptides. Food Chem 133:1349–1354CrossRefGoogle Scholar
  35. 35.
    Sharma S, Pathak N, Chattopadhyay K (2013) Osmolyte induced stabilization of protein molecules: a brief review. J Proteins Proteom 3:129–139Google Scholar
  36. 36.
    Kopcansky P, Siposova K, Melnikova L, Bednarikova Z, Timko M, Mitroova Z, Antosova A, Garamus VM, Petrenko VI, Avdeev MV, Gazova Z (2015) Destroying activity of magnetoferritin on lysozyme amyloid fibrils. J Magn Magn Mater 377:267–271CrossRefGoogle Scholar
  37. 37.
    Siposova K, Kubovcikova M, Bednarikova Z, Koneracka M, Zavisova V, Antosova A, Kopcansky P, Daxnerova Z, Gazova Z (2012) Depolymerization of insulin amyloid fibrils by albumin-modified magnetic fluid. Nanotechnology 23:055101CrossRefGoogle Scholar
  38. 38.
    Linse S, Cabaleiro-Lago C, Xue W, Lynch I, Lindman S, Thulin E, Radford SE, Dawson KA (2007) Nucleation of protein fibrillation by nanoparticles. Proc Natl Acad Sci USA 104: 8691–8696CrossRefGoogle Scholar
  39. 39.
    Rocha S, Thunemann AF, Pereira MC, Coelho M, Mohwald H, Brezesinski G (2008) Influence of fluorinated and hydrogenated nanoparticles on the structure and fibrillogenesis of amyloid beta-peptide. Biophys Chem 137:35–42CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Rupsa Chatterjee
    • 1
  • Vidyalatha Kolli
    • 1
  • Nandini Sarkar
    • 1
  1. 1.Department of Biotechnology and Medical EngineeringNational Institute of Technology RourkelaRourkelaIndia

Personalised recommendations