Identification of a new serine protease from polychaeta, Marphysa sanguinea, for its thrombolytic and anticoagulant activity

Seung Ju Yeon; Kyou Hee Shim; Jae Sang Hong; Hwa Sung Shin

Korean Journal of Chemical Engineering, Vol.34, No.3, 781-786, March, 2017

DOI10.1007/s11814-016-0331-z Export Citation

Identification of a new serine protease from polychaeta, Marphysa sanguinea, for its thrombolytic and anticoagulant activity

Seung Ju Yeon¹, Kyou Hee Shim¹, Jae Sang Hong^{2, 3}, and Hwa Sung Shin^{1, †}

¹Department of Biological Engineering, Inha University, Incheon 22212, Korea
²Department of Oceanography, Inha University, Incheon 22212, Korea
³Korea Institute of Coastal Ecology, Inc., Ojeong-gu, Bucheon 14449, Korea

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A serine protease was purified from Marphysa sanguinea through ammonium sulfate followed by ion exchange chromatography, and its N-terminal amino sequence was identified to be IVGGSEATPYQFPFQ. Fibrinolytic activity was depended on both direct fibrinolysis and indirect plasminogen-mediated cascade and had a consistent activity irrespective of pH. The serine protease could be confirmed to degrade α-, β-, and γ-chains of human fibrinogen through fibrinogenolytic assay and did not express significant cytotoxicity to endothelial cells. These imply the enzyme has anticoagulant as well as thrombolytic activity, not significantly impairing endothelial cells comprising brain blood brain barrier (BBB) tissue. Conclusively, the new serine protease is worthy of being a candidate to substitute tissue-Plasminogen Activator (t-PA) for acute ischemic reperfusion injury of brain.

Keywords:Serine Protease;Marphysa sanguinea;Brain Ischemia;Thrombolysis;Anticoagulant

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[2] Ingall T, J. Insur. Med., 36, 143 (2004)

[3] Thorvaldsen P, Asplund K, Kuulasmaa K, Rajakangas AM, Schroll M, Stroke, 26, 1 (1995)

[4] Simoons ML, Jaegere P, Domburg R, Boersma E, Maggioni AP, Franzosi MG, Leimberger JD, Califf R, Schroderm R, Knatterud G, Braunwald E, Lancet, 342, 1523 (1993)

[5] Fromm RE, Hoskins E, Cronin L, Pratt CM, Spencer WH, Roberts R, Ann. Emerg. Med., 20, 892 (1991)

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[7] Eisenberg PR, Sherman LA, Tiefenbrunn AJ, Ludbrook PA, Sobel BE, Jaffe AS, Thromb. Haemost., 57, 35 (1987)

[8] Nilsson T, Wallen P, Mellbring G, Eur. J. Haematol., 33, 49 (1984)

[9] Tsikouris JP, Tsikouris AP, Pharmacotherapy, 21, 207 (2001)

[10] Matsuo O, Rijken DC, Collen D, Thromb. Haemost., 45, 225 (1981)

[11] Hacke W, Albers G, Ai-Rawi Y, Bogousslavsky J, Davalos A, Eliasziw M, Fischer M, Furlan A, Kaste M, Lees KR, Soehgen M, Warach S, Stroke, 36, 1 (2005)

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[15] Matys T, Strickland S, Nat. Med., 9, 371 (2003)

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[19] Benchenane K, Lopez-Atalaya JP, Fernandez-Monreal M, Touzani O, Vivien D, Trends Neurosci., 27, 155 (2004)

[20] Kaur J, Zhao Z, Klein GM, Lo EH, Buchan AM, J. Cereb. Blood. Flow. Metab., 24, 945 (2004)

[21] Deng ZH, Wang SH, Li Q, Ji X, Zhang LZ, Hong M, Bioresour. Technol., 101(6), 1954 (2010)

[22] Zhang Y, Cui J, Zhang R, Wang Y, Hong M, Biochimie, 89, 93 (2007)

[23] Matsubara K, Hori K, Matsuura Y, Miyazawa K, Comp. Biochem. Physiol. B, Biochem. Mol. Biol., 125, 137 (2000)

[24] Wang S, Deng Z, Li Q, Ge X, Bo Q, Liu J, Cui J, Jiang X, Liu J, Zhang L, Hong M, Comp. Biochem. Physiol. B, Biochem. Mol. Biol., 159, 18 (2011)

[25] Collen D, Thromb. Haemost., 82, 259 (1999)

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[27] Camiolo SM, Thorsen S, Astrup T, Exp. Biol. Med., 138, 227 (1971)

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[29] James GT, Anal. Biochem., 86, 574 (1978)

[30] Choi JH, Sapkota K, Kim S, Kim SJ, Biochimie, 105, 45 (2014)

[31] Camiolo SM, Thorsen S, Astrup T, Exp. Biol. Med., 138, 277 (1971)

[32] Isaev NK, Stelmashook EV, Plotnikov EY, Khryapenkova TG, Lozier ER, Doludin YV, Silachev DN, Zorov DB, Biochem., 73, 1171 (2008)

[33] Alexandrov AV, Grotta JC, Neurology, 59, 862 (2002)

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[35] Farrell DH, Thiagarajan P, Chung DW, Davie EW, Proc. Natl. Acad. Sci. U. S. A., 89, 10729 (1992)

[36] Ouyang C, Huang TF, Biochim. Biophys. Acta - Enzymology, 571, 270 (1979)

[37] Lee JW, Seu JH, Rhee IK, Jin I, Kawamura Y, Park W, Biochem. Biophys. Res. Commun., 260, 665 (1999)

[38] Park JW, Park JE, Choi HK, Jung TW, Yoon SM, Lee JS, Process Biochem., 48(5-6), 979 (2013)