Flaxseed Cysteine Protease Exhibits Strong Anticoagulant, Antiplatelet, and Clot-Dissolving Properties

Autor:	Jayanna Kengaiah, Sharath Kumar M. Nandish, S M Santhosh, Ch Ramachandraiah, Chandramma, Devaraja Sannaningaiah, Ashwini Shivaiah, Thirunavukkarasu
Rok vydání:	2020
Předmět:	Platelet Aggregation Iodoacetic acid Hemorrhage Fibrinogen Hemolysis Biochemistry Fibrin Mice 03 medical and health sciences chemistry.chemical_compound Thrombin Cysteine Proteases Flax Casein medicine Animals Edema Humans Platelet Blood Coagulation 0303 health sciences Chromatography biology 030302 biochemistry & molecular biology Anticoagulants General Medicine Cysteine protease chemistry Clotting time biology.protein medicine.drug
Zdroj:	Biochemistry (Moscow). 85:1113-1126
ISSN:	1608-3040 0006-2979
Popis:	In this study, we purified and characterized flaxseed cysteine protease (FSCP) with strong anticoagulant, antiplatelet, and clot-dissolving properties. The enzyme was purified to homogeneity by a combination of gel permeation and ion-exchange column chromatography techniques. The purity of the enzyme was evaluated by SDS-PAGE, RP-HPLC, and MALDI-TOF. FSCP was observed as a single band of approximately 160 kDa in SDS-PAGE under reducing and non-reducing conditions. The exact molecular mass of FSCP was found to be 168 kDa by MALDI-TOF spectrometry. The CD spectra of FSCP revealed the presence of 25.6% helices, 25.8% turns, and 48% random coils with no beta-sheet structures. FSCP hydrolyzed both casein and gelatin with a specific activity of 3.5 and 4.2 unit/mg min respectively. The proteolytic activity of FSCP was completely abolished by iodoacetic acid (IAA), suggesting FSCP is a cysteine protease. The pH optimum for the proteolytic activity of FSCP was pH 6.0; the temperature optimum was 30°C. FSCP exhibited strong anticoagulant effect in both platelet-rich plasma (PRP) and platelet-poor plasma (PPP) by extending the clotting time from 222 to 1100 s and from 256 to 1210 s, respectively. FSCP degraded human fibrinogen and fibrin clots. The products of fibrinogen degradation by thrombin and FSCP were different. Furthermore, FSCP inhibited aggregation of washed platelets triggered by ADP, epinephrine, thrombin, collagen, arachidonic acid, and platelet activating factor (PAF). FSCP was found to be nontoxic as it did not damage the membrane of red blood cells (RBCs) and did not induce hemorrhage and edema in experimental mice.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::68d93f9a43f09a805801a447d13d7085 https://doi.org/10.1134/s0006297920090102 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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