Direct acting anti-thrombotic serine protease from brown seaweed Costaria costata

作者：Kim Dae Won; Sapkota Kumar; Choi Jun Hui; Kim Yoon Sik; Kim Seung; Kim Sung Jun^*

来源：Process Biochemistry, 2013, 48(2): 340-350.

DOI：10.1016/j.procbio.2012.12.012

• Algal protease
• Costaria costata
• Fibrino(geno)lysis
• Thrombosis

A direct acting antithrombotic serine protease (CCP) was purified from brown seaweed Costaria costata. CCP was a monomeric protease with molecular mass of 60,547.598 daltons as determined by mass spectrometry. The N-terminal sequence of CCP was SCNSCLDKVDADGLN. Proteolytic activity was inhibited by PMSF and APMSF. CCP exhibited high amidolytic activity toward substrate S-2251 with-apparent K-m and V-max values were 14.5 mu M and 183.5 U/ml respectively. Fibrin plate and fibrin zymography results revealed that CCP was able to degrade fibrin clots directly. It specifically hydrolyzed A alpha and alpha and B beta and beta chains followed by g and gamma-gamma chains of human fibrinogen and fibrin respectively. Cleavage of fibrin clot and fibrinogen was emphasized by observing the alteration of secondary structure using FTIR spectroscopy. Morphological alteration of fibrin clot was also evidenced by fluorescent microscopic observation. CCP reduced thrombus effectively in vitro. In vivo observation showed that it prevented/decreased thrombus formation in carrageenan-induced mice tail model. CCP prolonged activated partial thromboplastin time (APTT) and had little effect on prothrombin time (PT). Platelet function analyzer (PFA-100) tests showed that CCP prolonged closure time (CT). These data suggest that CCP could have therapeutic potential for the treatment of thrombosis.

• 出版日期2013-2