بررسی اثر مهاری تاکسیفولین بر فرایند فیبریلاسیون پروتئین لیزوزیم سفیده تخم مرغ

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشکده علوم زیستی، دانشگاه تحصیلات تکمیلی علوم پایه زنجان، زنجان، ایران

2 دانشکده علوم زیستی، دانشگاه تحصیلات تکمیلی علوم پایه زنجان، جاده گاوازنگ، زنجان، ایران

3 گروه زیست شناسی، واحد رودهن، دانشگاه آزاد اسلامی، رودهن، ایران

چکیده

در بین روشهای درمانی مورد استفاده برای بیماریهای مرتبط با تجمعات آمیلوئیدی، روشهای مبتنی بر استفاده از ترکیبات طبیعی با خاصیت آنتی آمیلوئیدوژنیک بسیار مورد توجه قرار گرفته است. اگرچه مطالعات in vitro و in vivo زیادی دلالت بر توانایی مهار فیبریلاسیون توسط این ترکیبات دارد ولی مکانیسم عمل این ترکیبات با یکدیگر متفاوت میباشد. در این مطالعه، با استفاده از رنج وسیعی از تکنیکها شامل فلورسانس تیوفلاوین T و نایل رد، جذب قرمز کنگو، و میکروسکوپ نیروی اتمی توانایی ملکول طبیعی تاکسیفولین در مهار فرایند فیبریلاسیون پروتئین لیزوزیم سفیده ی تخم مرغ مورد بررسی قرار گرفت. نتایج حاصل نشان داد که تاکسیفولین از طریق اتصال به تجمعات پیش فیبریلی اثرات مهاری خود را اعمال می نماید. بعلاوه، اتصال تاکسیفولین باعث تغییر مسیر فیبریلاسیون به سمت ایجاد ساختارهای کروی زنجیره ای شکل بزرگ میشود که دارای محتوای صفحات بتا کم و سطوح آبگریز در دسترس اندک میگردد. همچنین سنجش میزان فلورسانس تیوفلاوین T نشان داد که توانایی اتصال تاکسیفولین به تجمعات پیش فیبریلی با رشد آنها و تشکیل تجمعات پروتوفیبریلی بزرگ در انتهای فاز رشد کاهش می یابد. به نظر میرسد که نتایج حاصل در طراحی مهارکننده های تجمع پروتئینی مرتبط با بیماریهای تحلیل برنده ی سیستم عصبی موثر باشد.

کلیدواژه‌ها


عنوان مقاله [English]

A study on the inhibitory effects of taxifolin on amyloid fibrillation of hen egg white lysozyme

نویسندگان [English]

  • Mohsen Mahdavimehr 1
  • Ali Akbar Meratan 2
  • Ali Asghar Bagheri 3
  • Fariba Khosravinejad 3
1 Department of Biological Sciences, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan, Iran.
2 Department of Biological Sciences, Institute of Advanced Studies in Basic Sciences (IASBS), Zanjan, Iran.
3 Department of Biology, Islamic Azad University, Roudehen Branch,Roudehen,Iran
چکیده [English]

Among therapeutic approaches for amyloid-related diseases, attention has recently turned to the use of natural products as effective anti-aggregation compounds. Although a wealth of in vitro and in vivo evidence indicates some common inhibitory activity of these compounds, they don’t generally suggest the same mechanism of action. In the present study, using a range of techniques including Thioflavin T and Nile red fluorescence assays, Congo red absorbance measurements, and atomic force microscopy the ability of taxifolin on the inhibition of HEWL amyloid fibrillation was investigated. Obtained results demonstrated that taxifolin exerts its inhibitory effect by binding to HEWL prefibrillar species. Furthermore, it’s binding results in diverting the amyloid pathway toward formation of very large globular, chain-like aggregates with low β-sheet content and reduced solvent-exposed hydrophobic patches. ThT fluorescence measurements show that the binding capacity of taxifolin is significantly reduced, upon generation of large protofibrillar aggregates at the end of growth phase. We believe these results may help design promising inhibitors of protein aggregation for amyloid-related diseases.

کلیدواژه‌ها [English]

  • amyloid
  • Atomic Force Microscopy
  • Protein Aggregation
  • Taxifolin

[1].   Azami-Movahed M., Shariatizi S., Sabbaghian M., Ghasemi A., Ebrahim-HabibiA., Nemat-Gorgani M. 2013, Heme binding site in apomyoglobin may be effectively targeted with small molecules to control aggregation. International Journal of Biochemistry and Cell Biology. 45(2): 299-307.

[2].   Bartolini M., Andrisano V. 2010, Strategies for the inhibition of protein aggregation in human diseases. ChemBioChem. 11(8): 1018–1035.

[3].   Booth A.N., Deeds F. 1958, The toxicity and metabolism of dihydroquercetin. Journal of American Pharmaceutical Association. Americal Pharmaceutical Association. 47(3, part1):183–184.

[4].   Bouma B., Kroon-Batenburg L.M., Wu Y.P., Brünjes B., Posthuma G., Kranenburg O., de Groot P.G., Voest E.E., Gebbink M.F. 2003, Glycation induces formation of amyloid cross-beta structure in albumin. Journal of Biological Chemistry. 278 (43): 41810-9.

[5].   Bucciantini, M., Giannoni, E., Chiti, F., Baroni, F., Formigli, L., Zurdo, J., Taddei, N., Ramponi, G., Dobson, C.M., Stefani, M. 2002, Inherent toxicity of aggregates implies a common mechanism for protein misfolding diseases. Nature, 416(6880): 507–511.6.

[6].   Chiti F., Dobson C.M. 2006, Protein misfolding, functional amyloid and human disease. Annual Review of Biochemistry.75:333–366.

[7].   De Felice F.G., Ferreira S.T. 2002, β-Amyloid production, aggregation, and clearance as targets for therapy in Alzheimer’s disease. Cellular and Molecular Neurobiology. 22(5-6):545–5638.

[8].   Gazova Z., Siposova K., Kurin E., Mučaji P., Nagy M. 2013, Amyloid aggregation of lysozyme: the synergy study of red wine polyphenols. Proteins.81(6):994–1004.

[9].   Ghobeh M., Ahmadian S., Meratan A.A., Ebrahim-Habibi A., Ghasemi A., Shafizadeh M., Nemat-Gorgani M.2014, Interaction of Aβ(25–35) fibrillation products with mitochondria: effect of small-molecule natural products. Biopolymers. 102 (6):473–486.

[10].Goldberg M.E., Rudolph R., Jaenicke R. 1991, A kinetic study of the competition between renaturation and aggregation during the refolding of denatured-reduced egg white lysozyme. Biochemistry. 30(11): 2790–2797.

[11].Klunk W.E., Pettegrew J.W., Abraham D.J. 1989, Quantitative evaluation of Congo red binding to amyloid-like proteins with a beta-pleated sheet conformation. The Journal of Histochemistry and Cytochemistry. 37(8): 1273–1281.

[12].Krebs M.R., Wilkins D.K., Chung E.W., Pitkeathly M.C., Chamberlain A.K., Zurdo J., Robinson C.V., Dobson C.M.2000, Formation and seeding of amyloid fibrils from wild-type hen lysozyme and a peptide fragment from the β-domain. Journal of Molecular Biology. 300(3):541–549.

[13].Masuda M., Suzuki N., Taniguchi S., Oikawa T., Nonaka T., Iwatsubo T., Hisanaga S., Goedert M., Hasegawa M. 2006, Small molecule inhibitorsof alpha-synuclein filament assembly. Biochemistry. 45(19):6085–6094.

[14].Mishra, R., Sellin, D., Radovan, D., Gohlke, A., Winter, R. 2009, Inhibiting islet amyloid polypeptide fibril formation by the red wine compound resveratrol. ChemBioChem. 10(3): 445–449.

[15].Mishra, R., Sjölander, D., Hammarström, P. 2011, Spectroscopic characterization of diverse amyloid fibrils in vitro by the fluorescent dye Nile red. Molecular Biosystems. 7(4): 1232-40.

[16].Meratan A.A., Ghasemi A., Nemat-Gorgani M. 2011, Membrane integrity and amyloid cytotoxicity: a model study involving mitochondria and lysozyme fibrillation products. Journal of Molecular Biology. 409(5): 826–838.

[17].Pepys M.B., Hawking P.N., Booth D.R., Vigushin D.M., Tennent G.A., Soutar A.K., Totty N., Nguyen O., Blake C.C., Terry C.J., Feest T.G., Zalin A.M., Hsuan J.J. 1993, Human lysozyme gene mutations cause hereditary systemic amyloidosis. Nature. 362(6420): 553–557.

[18].Porat Y., Abramowitz A., Gazit E. 2006, Inhibition of amyloid fibril formation by polyphenols: structural similarity and aromatic interactions as a common inhibition mechanism. Chemical Biology & Drug Design. 67(1):27–37.

[19].Saito S., Yamamoto Y., Maki T., Hattori Y., Ito H., Mizuno K., Harada-Shiba M., Kalaria R.N., Fukushima M., Takahashi R., Ihara M.2017, Taxifolin inhibits amyloid-β oligomer formation and fully restores vascular integrity and memory in cerebral amyloid angiopathy. Acta Neuropathologica Communications. 5(1):26.

[20].Sato M., Murakami K., Uno M., Ikubo H., Nakagawa Y., Katayama S., Akagi K., Irie K.2013, Structure–Activity relationship for (‏)-taxifolin isolated from Silymarin as an inhibitor of Amyloid β Aggregation. Bioscience, Biotechnology, and Biochemistry. 77(5):1100–1103.

[21].Sato M., Murakami K., Uno M., Nakagawa Y., Katayama S., Akagi K., Masuda Y., Takegoshi K., Irie K.2013, Site-specific inhibitory mechanism for Amyloid β42 aggregation by catechol-type flavonoids targeting the Lys residues. Journal of Biological Chemistry. 288(32):23212–23224.

[22].Shariatizi S., Meratan A.A., Ghasemi A., Nemat-Gorgani M. 2015, Inhibition of amyloid fibrillation and cytotoxicity of lysozyme fibrillation products by polyphenols. International Journal of Biological Macromolecules. 80:95–106.

[23].Siebert K.J., Troukhanova N.V., Lynn, P.Y. 1996, Nature of polyphenol-protein interactions. Journal of Agricaltural and Food Chemistry. 44(1):80–85.

[24].Soares S., Mateus N., Freitas Vd. 2007, Interaction of different polyphenols with bovine serum albumin (BSA) and human salivary alpha-amylase (HSA) by fluorescence quenching. Journal of Agricaltural and Food Chemistry. 2007; 55(16):6726–6735.

[25].Soto C. 1999, Plaque busters: strategies to inhibit amyloid formation in Alzheimer’s disease. Molecular Medicine Today. 5(8):343–350.

[26].Stefani M., Rigacci S. 2013, Protein folding and aggregation into amyloid: the interference by natural phenolic compounds. International Journal of Molecular Sciences. 14(6):12411–12457.

[27].Sun X., Chen R.C., Yang Z.H., Sun G.B., Wang M., Ma X.J., Yang L.J., Sun X.B.2014, Taxifolin prevents diabetic cardiomyopathy in vivo and in vitro by inhibition of oxidative stress and cell apoptosis. Food and Chemical Toxicology. 63:221–232.

[28].Swaminathan R., Ravi V.K., Kumar S., Kumar M.V., Chandra N. 2011, Lysozyme: a model protein for amyloid research. Advances in Protein Chemistry and Structural Biology.84:63–111.

[29].Taniguchi S., Suzuki N., Masuda M., Hisanaga S., Iwatsubo T., Goedert M., Hasegawa M.2005, Inhibition of heparin-induced tau filament formation by phenothiazines, polyphenols, and porphyrins. Journal of Biological Chemistry. 280(9):7614–7623.

[30].Wang S.S.S., Chen P.H., Hung Y.T. 2006, Effects of p-benzoquinone and melatonin on amyloid fibrillogenesis of hen egg-white lysozyme. Journal of Molecular Catalysis B-Enzymatic. 43(1-4):49–57.

[31].Wang J.B., Wang Y.M., Zeng C.M. 2011, Quercetin inhibits amyloid fibrillation of bovine insulin and destabilizes preformed fibrils. Biochemical and Biophysical Research Communications. 415(4):675–679.

[32].Weidmann A.E. 2012, Dihydroquercetin: More than just an impurity? European Journal of Pharmacology. 684(1-3):19–26.

[33].Yang P., Xu F., Li H.F., Wang Y., Li F.C., Shang M.Y., Liu G.X., Wang X., Cai S.Q.2016, Detection of 191 Taxifolin metabolites and their distribution in Rats using HPLC-ESI-IT-TOF-MSn. Molecules. 21(9):1209.