DOI QR코드

DOI QR Code

Purification and characterization of β-secretase inhibitory peptide from sea hare (Aplysia kurodai) by enzymatic hydrolysis

  • Lee, Jung Kwon (Department of Marine Biotechnology, Gangneung-Wonju National University) ;
  • Kim, Sung Rae (Department of Marine Biotechnology, Gangneung-Wonju National University) ;
  • Byun, Hee-Guk (Department of Marine Biotechnology, Gangneung-Wonju National University)
  • Received : 2018.01.04
  • Accepted : 2018.02.13
  • Published : 2018.05.31

Abstract

Amyloid plaque, also called senile plaque, the product of aggregation of ${\beta}$-amyloid peptides ($A{\beta}$), is observed in brains of the patients with Alzheimer's disease (AD) and is one of the key factors in etiology of the disease. In this study, hydrolysates obtained from the sea hare (Aplysia kurodai) were investigated for ${\beta}$-secretase inhibitory peptide. The sea hare's muscle protein was hydrolyzed using six enzymes in a batch reactor. Trypsin hydrolysate had highest ${\beta}$-secretase inhibitory activity compared to the other hydrolysates. ${\beta}$-secretase inhibitory peptide was separated using Sephadex G-25 column chromatography and high-performance liquid chromatography on a C18 column. ${\beta}$-secretase inhibitory peptide was identified as eight amino acid residues of Val-Ala-Ala-Leu-Met-Leu-Phe-Asn by N-terminal amino acid sequence analysis. $IC_{50}$ value of purified ${\beta}$-secretase inhibitory peptide was $74.25{\mu}M$, and Lineweaver-Burk plots suggested that the peptide purified from sea hare muscle protein acts as a competitive inhibitor against ${\beta}$-secretase. Results of this study suggest that peptides derived from sea hare muscle may be beneficial as anti-dementia compounds in functional foods or as pharmaceuticals.

Keywords

References

  1. Association of Official Analytical Chemist (AOAC). Official Methods of Analysis. 17th ed. Washington: Association of Official Analytical Chemist; 2000.
  2. Choi YJ, Han YS. Protein and amino acid compositions in echiurid and sea hare muscles. Korean Soc Fisheries Aquatic Sci. 1985;18:550-6.
  3. Citron M. ${\beta}$-secretase as a target for the treatment of Alzheimer's disease. J Neurosci Res. 2002;70:373-9. https://doi.org/10.1002/jnr.10393
  4. Congreve M, Aharony D, Albert J, Callaghan O, Campbell J, Carr RA, Chessari G, Cowan S, Edwards PD, Frederickson M, McMenamin R, Murray CW, Patel S, Wallis N. Application of fragment screening by X-ray crystallography to the discovery of aminopyridines as inhibitors of beta-secretase. J Med Chem. 2007;50:1124-32. https://doi.org/10.1021/jm061197u
  5. Cook DG, Forman MS, Sung JC, Leight S, Kolson DL, Iwatsubo T, Lee VM, Doms RW. Alzheimer's a beta (1-42) is generated in the endoplasmic reticulum/intermediate compartment of NT2N cells. Nat Med. 1997;3:1021-3. https://doi.org/10.1038/nm0997-1021
  6. Dorrel S. Untangling Alzheimer's disease with ${\beta}$-secretase inhibitors. Drug Discov Today. 2000;5:316-7. https://doi.org/10.1016/S1359-6446(00)01531-2
  7. Duncan DB. Multiple range and multiple F-tests. Biometrics. 1955;11:1-12. https://doi.org/10.2307/3001478
  8. Garino C, Pietrancosta N, Laras Y, Moret V, Rolland A, Quelever G. BACE1 inhibitory activities of new substituted phenylpiperazine coupled to various heterocycles: chromene, coumarin and quinoline. Bioorg Med Chem Lett. 2006;16:1995-9. https://doi.org/10.1016/j.bmcl.2005.12.064
  9. Ghosh AK, Shin D, Koelsch G, Lin X, Ermolieff J, Tang J. Design of potent inhibitors for human brain memapsin 2 (${\beta}$-secretase). J Am Chem Soc. 2000;122:3522-3. https://doi.org/10.1021/ja000300g
  10. Hardy J, Allsop D. Amyloid deposition as the central event in the aetiology of Alzheimer's disease. Trends Pharmacol Sci. 1991;12:383-8. https://doi.org/10.1016/0165-6147(91)90609-V
  11. Hong L, He X, Huang X, Chang W, Tang J. Structural features of human memapsin 2 (${\beta}$-secretase) and their biological and pathological implications. Acta Biochim Biophys Sin. 2005;36:787-92.
  12. Huse JT, Liu K, Pijak DS, Carlin D, Lee VM, Doms RW. Beta-secretase processing in the trans-Golgi network preferentially generates truncated amyloid species that accumulate in Alzheimer's disease brain. J Biol Chem. 2002;277:16278-84. https://doi.org/10.1074/jbc.M111141200
  13. Je JY, Park PJ, Byun HG, Jung WK, Kim SK. Angiotensin I converting enzyme (ACE) inhibitory peptide derived from the sauce of fermented blue mussel, Mytilus edulis. Bioresour Technol. 2005;96:1624-9. https://doi.org/10.1016/j.biortech.2005.01.001
  14. Johnston JA, Liu WW, Coulson DTR, Todd S, Murphy S, Brennan S. Platelet ${\beta}$-secretase activity is increased in Alzheimer' disease. Neurobiol Aging. 2008;29:661-8. https://doi.org/10.1016/j.neurobiolaging.2006.11.003
  15. Kim WS. The study on anticancer activity from sea hare (Aplysia kurodai) eggs. Unpublished doctoral dissertation. Jeju: Jeju National University; 2008.
  16. Kimura T, Hamada Y, Stochaj M, Ikari H, Nagamine A, Abdel-Rahman H. Design and synthesis of potent ${\beta}$-secretase (BACE1) inhibitors with carboxylic acid bioisosteres. Bioorg Med Chem Lett. 2006;16:2380-6. https://doi.org/10.1016/j.bmcl.2006.01.108
  17. Korhonen H, Pihlanto A. Bioactive peptides: production and functionality. Int Dairy J. 2006;16:945-60. https://doi.org/10.1016/j.idairyj.2005.10.012
  18. Korhonen M, Pihlanto-Leppala A, Tupasela T. Impact of processing on bioactive proteins and peptides. Trends Food Sci Technol. 1998;9:307-19. https://doi.org/10.1016/S0924-2244(98)00054-5
  19. Kumagai Y, Ojima T. Isolation and characterization of two types of ${\beta}$-1,3-glucanases from the common sea hare Aplysia kurodai. Comp Biochem Physiol B. 2010;155:138-44. https://doi.org/10.1016/j.cbpb.2009.10.013
  20. Lee JK, Li-Chan ECY, Byun HG. Characterization of ${\beta}$-secretase inhibitory peptide purified from skate skin protein hydrolysate. Eur Food Res Technol. 2015;240:129-36. https://doi.org/10.1007/s00217-014-2314-9
  21. Lee JY, Lee SA, Kim JK, Chae CB, Kim Y. Interaction models of substrate peptides and ${\beta}$-secretase studied by NMR spectroscopy and molecular dynamics simulation. Mol Cells. 2009;27;651-6. https://doi.org/10.1007/s10059-009-0086-z
  22. Li-Chan ECY, Cheung IWY, Byun HG. Shrimp (Pandalopsis dispar) waste hydrolysate as a source of novel ${\beta}$-secretase inhibitors. Fish Aquat Sci. 2016;19:11. https://doi.org/10.1186/s41240-016-0008-x
  23. Menting KW, Claassen JA. ${\beta}$-secretase inhibitor; a promising novel therapeutic drug in Alzheimer's disease. Front Aging Neurosci. 2014;6:165.
  24. Neurath H, Walsh KA. Role of proteolytic enzymes in biological regulation. Proc Natl Acad Sci. 1976;73:3825-32. https://doi.org/10.1073/pnas.73.11.3825
  25. Park SH, Choung SY, Choi YJ. Immune regulating effect of polysaccharide fraction from sea hare (Aplysia kurodai). J Korean soc. Food Sci Nutr. 2011;40:372-8. https://doi.org/10.3746/jkfn.2011.40.3.372
  26. Shin MO. The antioxidative and antimicrobial effect of internal organs of Aplysia kurodai fraction. J Korean Fisheries Aquatic Sci. 2010;18:550-6.
  27. Silvestrelli G, Lanari A, Parnetti L, Tomassoni D, Amenta F. Treatment of Alzheimer's disease: from pharmacology to a better understanding of disease pathophysiology. Mech Ageing Dev. 2006;127:148-57. https://doi.org/10.1016/j.mad.2005.09.018
  28. Stroud RM, Kossiakoff AA, Chambers JL. Mechanisms of zymogen activation. Annu Rev Biophys Bioeng. 1997;6:177-93.
  29. Turner RT, Koelsch G, Hong L, Castanheira P, Ermolieff J, Ghosh AK. Subsite specificity of memapsin 2 (${\beta}$-secretase): implications for inhibitor design. Biochemist. 2001;40:10001-6. https://doi.org/10.1021/bi015546s
  30. Vassar R, Bennett BD, Babu-Khan S, Kahn S, Mendiaz EA, Denis P. ${\beta}$-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science. 1999;286:735-41. https://doi.org/10.1126/science.286.5440.735

Cited by

  1. A peptide fraction of Olive Flounder (Paralichthys olivaceus) Skin Hydrolysate Inhibits Amyloid-β Generation in SH-SY5Y Cells via Suppression of BACE1 Expression vol.27, pp.1, 2021, https://doi.org/10.1007/s10989-020-10113-8
  2. Marine-Derived Compounds with Anti-Alzheimer’s Disease Activities vol.19, pp.8, 2021, https://doi.org/10.3390/md19080410