Purification and Characterization of a Novel Fibrinolytic Metalloprotease from Nereis virens
DENG Zhi-Hui1)* , SUN He2) , LIN Yan1) , WANG Guo-Zhong1) , JIN Li1) , HONG Min3)
(1) Department of Pathophysiology, Qiqihar Medical University, Qiqihar 161006, China;
2) Department of Histoembryology, Qiqihar Medical University, Qiqihar 161006, China;
3) Department of Biochemistry and Molecular Biology, Norman Bethune College of Medicine,
Jilin University, Changchun 130021, China)
Abstract：A novel fibrinolytic metalloprotease from Nereis virens, named NVMP, was purified by using ammonium sulfate precipitation, anion exchange chromatography on DEAE-Sepharose FF, cation exchange chromatography on CM-Sepharose FF and gel-filtration chromatography on Sephacryl S-100 HR. NVMP consisted of a single polypeptide chain with a molecular weight of 28~32 kD, which was determined by SDS-PAGE and Matrix-assisted laser desorption/ionization time-of-flight mass spectrum (MALDI-TOF MS). The isoelectric point of NVMP determined by isoelectric focusing electrophoresis (IEF) was 8.0. NVMP activity was strongly inhibited by EGTA, indicating that it was a metalloprotease and the maximum activity of the enzyme was observed at 40 ℃ and pH 6.0. The enzymatic activity of this metalloprotease was inhibited by Cu2+, Co2+ and Zn2+, but enhanced by Ca2+ and Mg2+. The peptide mass fingerprinting (PMF) revealed that NVMP was a newly discovered protein. More importantly, NVMP degraded fibrin either directly or indirectly through generating plasmin from activation of plasminogen. NVMP may thus have a medical value for preventing and treating thrombosis.
邓志会, 孙贺, 林岩, 王国忠, 金莉, 洪敏. 一种新型具有纤溶活性的沙蚕金属蛋白酶的分离纯化及鉴定[J]. 中国生物化学与分子生物学报, 2011, 27(8): 768-774.
DENG Zhi-Hui, SUN He, LIN Yan, WANG Guo-Zhong, JIN Li, HONG Min. Purification and Characterization of a Novel Fibrinolytic Metalloprotease from Nereis virens. Chinese Journal of Biochemistry and Molecular Biol, 2011, 27(8): 768-774.
