title: Purification and partial characterization of a novel fibrinogenase from the venom of Deinagkistrodon acutus: inhibition of platelet aggregation.
abstract: A novel fibrinogenase, DAnase, was purified from the venom of Deinagkistrodon acutus by a combination of anion and cation exchange chromatography. Unlike other fibrinogenases which are usually single polypeptide chain proteins, the enzyme was a disulfide-linked dimer with an isoelectric point of 6.03 and an apparent molecular weight of 25kDa on SDS-polyacrylamide gel electrophoresis. DAnase showed α-fibrinogenase activity devoid of fibrinolytic activity. It hydrolyzed rapidly the Aα-chain of fibrinogen and followed by the Bβ-chain and did not cleave the γ-chain. It also exhibited arginine esterase activity. The fibrinogenolytic and arginine esterase activities were completely inhibited by phenylmethanesulfonyl fluoride or tris-(2-carboxyethyl)phosphine hydrochloride, but not by EDTA, indicating that DAnase is a serine protease requiring disulfide bridge(s) for its activity. The protease strongly inhibited ADP-induced platelet aggregation in human platelet-rich plasma but was lack of ADPase activity, indicating that its fibrinogenolytic activity is involved in its inhibition of ADP-induced platelet aggregation. DAnase was devoid of hemorrhagic activity and Factor XIII activation activity. DAnase may have a potential clinical application for the therapy of thrombosis disease.
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei 230026, PR China.
authors.forename: Shasha
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei 230026, PR China. Electronic address: xuxl@ustc.edu.cn.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei 230026, PR China.
authors.forename: Shang
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei 230026, PR China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei 230026, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Rong
authors.affiliation: Department of Chemistry, University of Science and Technology of China, Hefei 230026, PR China.
authors.forename: Bing
authors.initials: B
authors.lastname: Li
mesh_terms: D000818:Animals; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000302:isolation & purification / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects*; D055503:Protein Multimerization; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D057057:Serine Proteases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D017819:Viperidae
chemical_list: D003435:Crotalid Venoms; D005340:Fibrinogen; C000594252:DAnase alpha-fibrinogenase, Deinagkistrodon acutus; D057057:Serine Proteases; D012697:Serine Endopeptidases
keywords: DAnase; Deinagkistrodon acutus; Non-hemorrhagic fibrinogenase; Platelet aggregation; Serine protease; Snake venom