tailieunhanh - Báo cáo hóa học: " Purification and characterization of novel fibrinolytic proteases as potential antithrombotic agents from earthworm Perionyx excavatus"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí hóa hoc quốc tế đề tài : Purification and characterization of novel fibrinolytic proteases as potential antithrombotic agents from earthworm Perionyx excavatus | Phan et al. AMB Express 2011 1 26 http content 1 1 26 o AMB Express a SpringerOpen Journal ORIGINAL Open Access Purification and characterization of novel fibrinolytic proteases as potential antithrombotic agents from earthworm Perionyx excavatus Tram Thi Bich Phan1 Tien Duy Ta2 Dung Thi Xuan Nguyen3 Lambertus AM Van Den Broek4 and Giang Thi Huong Duong3 Abstract Six protease fractions namely FI FII FIII-1 FIII-2 FIII-3 and FIV were isolated from Perionyx excavatus earthworm biomass by acetone precipitation followed by serial chromatography using anion exchange hydrophobic interaction and size exclusion chromatography. All fractions exhibited strong hydrolytic activity towards casein. The activity of six fractions towards fibrin determined by fibrin plate assay ranged from 44 to 831 plasmin and ranked as FIII-3 FIII-2 FI FIII-1 FIV FII. Casein degradation was optimal at pH 7 and 11 and at 45-60 C. All fractions were considerably stable at high temperature and wide pH range. They were completely inhibited by phenylmethylsulfonyl fluoride PMSF . The molecular weights MW and isoelectric points pI determined by 2D-electrophoresis were kDa and respectively. Tandem mass spectrometry MS analysis was used to deduce the amino acid sequences of some peptides from FIII-1 and FIII-2. The sequences shared and similarity respectively with the fibrinolytic enzymes from two related earthworm species Lumbricus rubellus and Eisenia fetida. The P. excavatus proteases were classified as serine proteases. They could perform rapid hydrolysis on both coagulated fibrous fibrin and soluble fibrinogen monomers without the presence of activators such as tPA or urokinase. Keywords chromatography fibrinolysis Perionyx excavatus PMSF serine protease tandem MS analysis Introduction Cardiovascular diseases have become one of the biggest concerns all over the world Grundy et al. 1999 . Among these thrombosis is the most widespread within .