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.