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Journal of Solution Chemistry

, Volume 47, Issue 5, pp 939–949 | Cite as

Salt Modulated Fibrillar Aggregation of the Sweet Protein MNEI in Aqueous Solution

  • Federica Donnarumma
  • Alessandro Emendato
  • Serena Leone
  • Carmine Ercole
  • Gerardino D’Errico
  • Delia Picone
Article
  • 94 Downloads

Abstract

The mechanism of conversion of globular native proteins into amyloid fibrils represents one of the most attractive research topics in biophysics, because of its involvement in the development of severe pathologies and in various biotechnological processes. Aqueous medium properties, such as pH and ionic strength, as well as interactions with other species in solution, play a key role in tuning the fibrillization process. Here, we describe a comparative study of the influence of different ions from the Hofmeister series on the thermal unfolding and aggregation propensity of MNEI, a model protein, selected because of its tendency to form amyloid aggregates at acidic pH, even at temperatures well below its melting temperature. By selecting a temperature at which only negligible amounts of protein are unfolded, we have focused on the effect of ions on fibril formation. ThT fluorescence experiments indicated that all the salts examined increased the rate and the extent of fibrillization. Moreover, we found that anions, particularly sulfate, strongly influence the process, which instead is only marginally affected by different cations. Finally, a specific link to the chloride concentration was detected.

Keywords

Protein aggregation Protein–ions interactions Hofmeister series ThT fluorescence 

Notes

Acknowledgements

The financial support of the “Fondazione con il Sud” (Grant No. 2011-PDR-19) is gratefully acknowledged. FD was recipient a fellowship financed by Regione Campania (POR Campania FSE 2014–2020).

Supplementary material

10953_2018_764_MOESM1_ESM.docx (576 kb)
Supplementary material 1 (DOCX 575 kb)

References

  1. 1.
    Buell, A.K., Hung, P., Salvatella, X., Welland, M.E., Dobson, C.M., Knowles, T.P.J.: Electrostatic effects in filamentous protein aggregation. Biophys. J. 104, 1116–1126 (2013).  https://doi.org/10.1016/j.bpj.2013.01.031 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Marek, P.J., Patsalo, V., Green, D.F., Raleigh, D.P.: Ionic strength effects on amyloid formation by amylin are a complicated interplay among Debye screening, ion selectivity, and Hofmeister effects. Biochemistry 51, 8478–8490 (2012).  https://doi.org/10.1021/bi300574r CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Sirangelo, I., Iannuzzi, C.: The role of metal binding in the amyotrophic lateral sclerosis-related aggregation of copper–zinc superoxide dismutase. Molecules 22, 1429 (2017).  https://doi.org/10.3390/molecules22091429 CrossRefGoogle Scholar
  4. 4.
    Koelsch, P., Motschmann, H.: An experimental route to Hofmeister. Curr. Opin. Colloid Interface Sci. 9, 87–91 (2004).  https://doi.org/10.1016/j.cocis.2004.05.009 CrossRefGoogle Scholar
  5. 5.
    Lee, E., Choi, J.-H., Cho, M.: The effect of Hofmeister anions on water structure at protein surfaces. Phys. Chem. Chem. Phys. 19, 20008–20015 (2017).  https://doi.org/10.1039/C7CP02826A CrossRefPubMedGoogle Scholar
  6. 6.
    Násztor, Z., Dér, A., Bogár, F.: Ion-induced alterations of the local hydration environment elucidate Hofmeister effect in a simple classical model of Trp-cage miniprotein. J. Mol. Model. 23, 298 (2017).  https://doi.org/10.1007/s00894-017-3471-0 CrossRefPubMedGoogle Scholar
  7. 7.
    Bye, J.W., Baxter, N.J., Hounslow, A.M., Falconer, R.J., Williamson, M.P.: Molecular mechanism for the Hofmeister effect derived from NMR and DSC measurements on barnase. ACS Omega. 1, 669–679 (2016).  https://doi.org/10.1021/acsomega.6b00223 CrossRefGoogle Scholar
  8. 8.
    Imperatore, R., Vitiello, G., Ciccarelli, D., D’Errico, G.: Effects of salts on the micellization of a short-tailed nonionic ethoxylated surfactant: an intradiffusion study. J. Solution Chem. 43, 227–239 (2014).  https://doi.org/10.1007/s10953-014-0133-z CrossRefGoogle Scholar
  9. 9.
    Dér, A., Kelemen, L., Fábián, L., Taneva, S.G., Fodor, E., Páli, T., Cupane, A., Cacace, M.G., Ramsden, J.J.: Interfacial water structure controls protein conformation. J. Phys. Chem. B. 111, 5344–5350 (2007).  https://doi.org/10.1021/jp066206p CrossRefPubMedGoogle Scholar
  10. 10.
    Chaplin, M.F.: A proposal for the structuring of water. Biophys. Chem. 83, 211–221 (2000)CrossRefPubMedGoogle Scholar
  11. 11.
    Salis, A., Ninham, B.W.: Models and mechanisms of Hofmeister effects in electrolyte solutions, and colloid and protein systems revisited. Chem. Soc. Rev. 43, 7358–7377 (2014).  https://doi.org/10.1039/C4CS00144C CrossRefPubMedGoogle Scholar
  12. 12.
    Kim, S.-H., Kang, C.-H., Kim, R., Cho, J.M., Lee, Y.-B., Lee, T.-K.: Redesigning a sweet protein: increased stability and renaturability. Protein Eng. 2, 571–575 (1989).  https://doi.org/10.1093/protein/2.8.571 CrossRefPubMedGoogle Scholar
  13. 13.
    Tancredi, T., Iijima, H., Saviano, G., Amodeo, P., Temussi, P.A.: Structural determination of the active site of a sweet protein A 1H NMR investigation of pMNEI. FEBS Lett. 310, 27–30 (1992).  https://doi.org/10.1016/0014-5793(92)81138-C CrossRefPubMedGoogle Scholar
  14. 14.
    Esposito, V., Gallucci, R., Picone, D., Saviano, G., Tancredi, T., Temussi, P.A.: The importance of electrostatic potential in the interaction of sweet proteins with the sweet taste receptor. J. Mol. Biol. 360, 448–456 (2006).  https://doi.org/10.1016/j.jmb.2006.05.020 CrossRefPubMedGoogle Scholar
  15. 15.
    Leone, S., Picone, D.: Molecular dynamics driven design of pH-stabilized mutants of MNEI, a sweet protein. PLoS ONE 11, e0158372 (2016).  https://doi.org/10.1371/journal.pone.0158372 CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Leone, S., Pica, A., Merlino, A., Sannino, F., Temussi, P.A., Picone, D.: Sweeter and stronger: enhancing sweetness and stability of the single chain monellin MNEI through molecular design. Sci. Rep. 6, 34045 (2016).  https://doi.org/10.1038/srep34045 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Liu, Q., Li, L., Yang, L., Liu, T., Cai, C., Liu, B.: Modification of the sweetness and stability of sweet-tasting protein monellin by gene mutation and protein engineering. Biomed. Res. Int. 2016, e3647173 (2016).  https://doi.org/10.1155/2016/3647173 CrossRefGoogle Scholar
  18. 18.
    Cai, C., Li, L., Lu, N., Zheng, W., Yang, L., Liu, B.: Expression of a high sweetness and heat-resistant mutant of sweet-tasting protein, monellin, in Pichia pastoris with a constitutive GAPDH promoter and modified N-terminus. Biotechnol. Lett. (2016).  https://doi.org/10.1007/s10529-016-2182-4 CrossRefPubMedGoogle Scholar
  19. 19.
    Zheng, W., Yang, L., Cai, C., Ni, J., Liu, B.: Expression, purification and characterization of a novel double-sites mutant of the single-chain sweet-tasting protein monellin (MNEI) with both improved sweetness and stability. Protein Expr. Purif. 143, 52–56 (2018).  https://doi.org/10.1016/j.pep.2017.10.010 CrossRefPubMedGoogle Scholar
  20. 20.
    Morris, J.A., Cagan, R.H.: Purification of monellin, the sweet principle of Dioscoreophyllum cumminsii. Biochim. Biophys. Acta. 261, 114–122 (1972).  https://doi.org/10.1016/0304-4165(72)90320-0 CrossRefPubMedGoogle Scholar
  21. 21.
    Temussi, P.A.: Why are sweet proteins sweet? Interaction of brazzein, monellin and thaumatin with the T1R2–T1R3 receptor. FEBS Lett. 526, 1–4 (2002)CrossRefPubMedGoogle Scholar
  22. 22.
    Spadaccini, R., Crescenzi, O., Tancredi, T., De Casamassimi, N., Saviano, G., Scognamiglio, R., Di Donato, A., Temussi, P.A.: Solution structure of a sweet protein: NMR study of MNEI, a single chain monellin. J. Mol. Biol. 305, 505–514 (2001).  https://doi.org/10.1006/jmbi.2000.4304 CrossRefPubMedGoogle Scholar
  23. 23.
    Hobbs, J.R., Munger, S.D., Conn, G.L.: Monellin (MNEI) at 1.15 Å resolution. Acta Crystallogr. F 63, 162–167 (2007).  https://doi.org/10.1107/s1744309107005271 CrossRefGoogle Scholar
  24. 24.
    Ghiso, J., Jensson, O., Frangione, B.: Amyloid fibrils in hereditary cerebral hemorrhage with amyloidosis of Icelandic type is a variant of gamma-trace basic protein (cystatin C). Proc. Natl. Acad. Sci. USA 83, 2974–2978 (1986)CrossRefPubMedGoogle Scholar
  25. 25.
    Murzin, A.G.: Sweet-tasting protein monellin is related to the cystatin family of thiol proteinase inhibitors. J. Mol. Biol. 230, 689–694 (1993).  https://doi.org/10.1006/jmbi.1993.1186 CrossRefPubMedGoogle Scholar
  26. 26.
    Konno, T.: Multistep nucleus formation and a separate subunit contribution of the amyloidgenesis of heat-denatured monellin. Protein Sci. 10, 2093–2101 (2001).  https://doi.org/10.1110/ps.20201 CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Konno, T., Murata, K., Nagayama, K.: Amyloid-like aggregates of a plant protein: a case of a sweet-tasting protein, monellin. FEBS Lett. 454, 122–126 (1999).  https://doi.org/10.1016/S0014-5793(99)00789-9 CrossRefPubMedGoogle Scholar
  28. 28.
    Rega, M.F., Di Monaco, R., Leone, S., Donnarumma, F., Spadaccini, R., Cavella, S., Picone, D.: Design of sweet protein based sweeteners: Hints from structure–function relationships. Food Chem. 173, 1179–1186 (2015).  https://doi.org/10.1016/j.foodchem.2014.10.151 CrossRefPubMedGoogle Scholar
  29. 29.
    Esposito, V., Guglielmi, F., Martin, S.R., Pauwels, K., Pastore, A., Piccoli, R., Temussi, P.A.: Aggregation mechanisms of cystatins: a comparative study of monellin and oryzacystatin. Biochemistry 49, 2805–2810 (2010).  https://doi.org/10.1021/bi902039s CrossRefPubMedGoogle Scholar
  30. 30.
    Pica, A., Leone, S., Di, G., Donnarumma, F., Emendato, A., Rega, M.F., Merlino, A., Picone, D.: pH driven fibrillar aggregation of the super-sweet protein Y65R-MNEI: a step-by-step structural analysis. Biochim. Biophys. Acta 1862, 808–815 (2018).  https://doi.org/10.1016/j.bbagen.2017.12.012 CrossRefGoogle Scholar
  31. 31.
    Patra, A.K., Udgaonkar, J.B.: Characterization of the folding and unfolding reactions of single-chain monellin: evidence for multiple intermediates and competing pathways. Biochemistry 46, 11727–11743 (2007).  https://doi.org/10.1021/bi701142a CrossRefPubMedGoogle Scholar
  32. 32.
    Kimura, T., Maeda, A., Nishiguchi, S., Ishimori, K., Morishima, I., Konno, T., Goto, Y., Takahashi, S.: Dehydration of main-chain amides in the final folding step of single-chain monellin revealed by time-resolved infrared spectroscopy. Proc. Natl. Acad. Sci. USA 105, 13391–13396 (2008).  https://doi.org/10.1073/pnas.0801316105 CrossRefPubMedGoogle Scholar
  33. 33.
    Jha, S.K., Udgaonkar, J.B.: Direct evidence for a dry molten globule intermediate during the unfolding of a small protein. Proc. Natl. Acad. Sci. USA 106, 12289–12294 (2009).  https://doi.org/10.1073/pnas.0905744106 CrossRefPubMedGoogle Scholar
  34. 34.
    Malhotra, P., Udgaonkar, J.B.: High-energy intermediates in protein unfolding characterized by thiol labeling under native like conditions. Biochemistry 53, 3608–3620 (2014).  https://doi.org/10.1021/bi401493t CrossRefPubMedGoogle Scholar
  35. 35.
    Goluguri, R.R., Udgaonkar, J.B.: Rise of the helix from a collapsed globule during the folding of monellin. Biochemistry 54, 5356–5365 (2015).  https://doi.org/10.1021/acs.biochem.5b00730 CrossRefPubMedGoogle Scholar
  36. 36.
    Maity, H., Reddy, G.: Thermodynamics and kinetics of single-chain monellin folding with structural insights into specific collapse in the denatured state ensemble. J. Mol. Biol. (2017).  https://doi.org/10.1016/j.jmb.2017.09.009 CrossRefPubMedGoogle Scholar
  37. 37.
    Aghera, N., Dasgupta, I., Udgaonkar, J.B.: A buried ionizable residue destabilizes the native state and the transition state in the folding of monellin. Biochemistry 51, 9058–9066 (2012).  https://doi.org/10.1021/bi3008017 CrossRefPubMedGoogle Scholar
  38. 38.
    Spadaccini, R., Leone, S., Rega, M.F., Richter, C., Picone, D.: Influence of pH on the structure and stability of the sweet protein MNEI. FEBS Lett. 590, 3681–3689 (2016).  https://doi.org/10.1002/1873-3468.12437 CrossRefPubMedGoogle Scholar
  39. 39.
    Leone, S., Sannino, F., Tutino, M.L., Parrilli, E., Picone, D.: Acetate: friend or foe? Efficient production of a sweet protein in Escherichia coli BL21 using acetate as a carbon source. Microb. Cell Factories. 14, 106 (2015).  https://doi.org/10.1186/s12934-015-0299-0 CrossRefGoogle Scholar
  40. 40.
    Greenfield, N.J.: Using circular dichroism collected as a function of temperature to determine the thermodynamics of protein unfolding and binding interactions. Nat. Protoc. 1, 2527–2535 (2006).  https://doi.org/10.1038/nprot.2006.204 CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Xue, C., Lin, T.Y., Chang, D., Guo, Z.: Thioflavin T as an amyloid dye: fibril quantification, optimal concentration and effect on aggregation. R. Soc. Open Sci. 4, 160696 (2017).  https://doi.org/10.1098/rsos.160696 CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Sabate, R., Rodriguez-Santiago, L., Sodupe, M., Saupe, S.J., Ventura, S.: Thioflavin-T excimer formation upon interaction with amyloid fibers. Chem. Commun. 49, 5745–5747 (2013).  https://doi.org/10.1039/C3CC42040J CrossRefGoogle Scholar
  43. 43.
    Adrover, M., Mariño, L., Sanchis, P., Pauwels, K., Kraan, Y., Lebrun, P., Vilanova, B., Muñoz, F., Broersen, K., Donoso, J.: Mechanistic insights in glycation-induced protein aggregation. Biomacromolecules 15, 3449–3462 (2014).  https://doi.org/10.1021/bm501077j CrossRefPubMedGoogle Scholar
  44. 44.
    Mariño, L., Pauwels, K., Casasnovas, R., Sanchis, P., Vilanova, B., Muñoz, F., Donoso, J., Adrover, M.: Ortho-methylated 3-hydroxypyridines hinder hen egg-white lysozyme fibrillogenesis. Sci. Rep. (2015).  https://doi.org/10.1038/srep12052 CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Stewart, K.L., Radford, S.E.: Amyloid plaques beyond Aβ: a survey of the diverse modulators of amyloid aggregation. Biophys. Rev. 9, 405–419 (2017).  https://doi.org/10.1007/s12551-017-0271-9 CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Han, J.Y., Choi, T.S., Kim, H.I.: Molecular role of Ca2+ and hard divalent metal cations on accelerated fibrillation and interfibrillar aggregation of α-synuclein. Sci. Rep. 8, 1895 (2018).  https://doi.org/10.1038/s41598-018-20320-5 CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Ramis, R., Ortega-Castro, J., Vilanova, B., Adrover, M., Frau, J.: A systematic DFT study of some plausible Zn(II) and Al(III) interaction sites in N-terminally acetylated α-synuclein. J. Phys. Chem. A 122, 690–699 (2018).  https://doi.org/10.1021/acs.jpca.7b10744 CrossRefPubMedGoogle Scholar
  48. 48.
    Ramis, R., Ortega-Castro, J., Vilanova, B., Adrover, M., Frau, J.: Copper(II) binding sites in N-terminally acetylated α-synuclein: a theoretical rationalization. J. Phys. Chem. A 121, 5711–5719 (2017).  https://doi.org/10.1021/acs.jpca.7b03165 CrossRefPubMedGoogle Scholar
  49. 49.
    Klement, K., Wieligmann, K., Meinhardt, J., Hortschansky, P., Richter, W., Fändrich, M.: Effect of different salt ions on the propensity of aggregation and on the structure of Alzheimer’s Aβ(1-40) amyloid fibrils. J. Mol. Biol. 373, 1321–1333 (2007).  https://doi.org/10.1016/j.jmb.2007.08.068 CrossRefPubMedGoogle Scholar
  50. 50.
    Munishkina, L.A., Henriques, J., Uversky, V.N., Fink, A.L.: Role of protein–water interactions and electrostatics in alpha-synuclein fibril formation. Biochemistry 43, 3289–3300 (2004).  https://doi.org/10.1021/bi034938r CrossRefPubMedGoogle Scholar
  51. 51.
    Yeh, V., Broering, J.M., Romanyuk, A., Chen, B., Chernoff, Y.O., Bommarius, A.S.: The Hofmeister effect on amyloid formation using yeast prion protein. Protein Sci. 19, 47–56 (2010).  https://doi.org/10.1002/pro.281 CrossRefPubMedGoogle Scholar
  52. 52.
    Arosio, P., Jaquet, B., Wu, H., Morbidelli, M.: On the role of salt type and concentration on the stability behavior of a monoclonal antibody solution. Biophys. Chem. 168–169, 19–27 (2012).  https://doi.org/10.1016/j.bpc.2012.05.004 CrossRefPubMedGoogle Scholar
  53. 53.
    Yang, D.S., Yip, C.M., Huang, T.H., Chakrabartty, A., Fraser, P.E.: Manipulating the amyloid-beta aggregation pathway with chemical chaperones. J. Biol. Chem. 274, 32970–32974 (1999)CrossRefPubMedGoogle Scholar
  54. 54.
    Sikkink, L.A., Ramirez-Alvarado, M.: Salts enhance both protein stability and amyloid formation of an immunoglobulin light chain. Biophys. Chem. 135, 25–31 (2008).  https://doi.org/10.1016/j.bpc.2008.02.019 CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Zhang, Y., Cremer, P.S.: Chemistry of Hofmeister anions and osmolytes. Annu. Rev. Phys. Chem. 61, 63–83 (2010).  https://doi.org/10.1146/annurev.physchem.59.032607.093635 CrossRefPubMedGoogle Scholar
  56. 56.
    Light, T.P., Corbett, K.M., Metrick, M.A., MacDonald, G.: Hofmeister ion-induced changes in water structure correlate with changes in solvation of an aggregated protein complex. Langmuir 32, 1360–1369 (2016).  https://doi.org/10.1021/acs.langmuir.5b04489 CrossRefPubMedGoogle Scholar
  57. 57.
    Parsons, D.F., Boström, M., Nostro, P.L., Ninham, B.W.: Hofmeister effects: interplay of hydration, nonelectrostatic potentials, and ion size. Phys. Chem. Chem. Phys. 13, 12352–12367 (2011).  https://doi.org/10.1039/c1cp20538b CrossRefPubMedGoogle Scholar
  58. 58.
    Gokarn, Y.R., Fesinmeyer, R.M., Saluja, A., Razinkov, V., Chase, S.F., Laue, T.M., Brems, D.N.: Effective charge measurements reveal selective and preferential accumulation of anions, but not cations, at the protein surface in dilute salt solutions. Protein Sci. 20, 580–587 (2011).  https://doi.org/10.1002/pro.591 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Federica Donnarumma
    • 1
  • Alessandro Emendato
    • 1
  • Serena Leone
    • 1
  • Carmine Ercole
    • 1
  • Gerardino D’Errico
    • 1
  • Delia Picone
    • 1
  1. 1.Department of Chemical SciencesUniversity of Naples Federico II, Complesso Universitario di Monte Sant’AngeloNaplesItaly

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