title: Expression and characterization of a fibrinogenolytic enzyme from horsefly salivary gland.
abstract: Enzymes from various natural resources are valuable in management of thrombosis. Blood-sucking arthropods are one of these resources because they have a wide array of anti-hemostasis molecules in their salivary gland. However, it is difficult to purify enough protein samples from the salivary glands for pharmacological studies. In this work, a fibrinogenolytic enzyme (tablysin 2) identified from salivary glands of the horsefly Tabanus yao was expressed in Escherichia coli to further study its biological activities. The primary structure of tablysin 2 showed significant domain similarity to arthropod proteins from the antigen 5 family containing SCP domain, whose biological functions are poorly understood. Tablysin 2 cleaved the Aα and part of Bβ chains of fibrinogen and did not affect γ chain and fibrin. It inhibited platelet aggregation induced by ADP. It did not directly induce hemorrhage or activate plasminogen. The fibrinogenolytic activity of tablysin 2 provides a clue for the functions of antigen 5-related proteins in other haematophagous arthropods. This work demonstrate a method of expression of arthropod salivary proteins which are difficult to obtain from natural resources for further functional studies.
authors.affiliation: The Affiliated Stomatological Hospital of Kunming Medicinal College, Kunming, 650221, Yunnan, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Sheng
authors.affiliation: Key Laboratory of Animal Models and Human Disease Mechanism of the Chinese Academy of Science & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gao
authors.affiliation: Key Laboratory of Animal Models and Human Disease Mechanism of the Chinese Academy of Science & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; Graduate University of Chinese Academy of Sciences, Beijing, 100009, China.
authors.forename: Xiancui
authors.initials: X
authors.lastname: Lu
authors.affiliation: Key Laboratory of Animal Models and Human Disease Mechanism of the Chinese Academy of Science & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; Graduate University of Chinese Academy of Sciences, Beijing, 100009, China.
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Models and Human Disease Mechanism of the Chinese Academy of Science & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; Graduate University of Chinese Academy of Sciences, Beijing, 100009, China.
authors.forename: Yalan
authors.initials: Y
authors.lastname: Han
authors.affiliation: Key Laboratory of Animal Models and Human Disease Mechanism of the Chinese Academy of Science & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; United Laboratory of Natural Peptide of University of Science and Technology of China, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; Sino-African Joint Research Center, Chinese Academy of Science, Wuhan, Hubei, 430074, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Meng
authors.affiliation: Yunnan Clinical Research Center of Breast Cancer, The Third Affiliated Hospital of Kunming Medical College, Kunming, 650032, China. Electronic address: chenwenlin@aliyun.com.
authors.forename: Wenlin
authors.initials: W
authors.lastname: Chen
authors.affiliation: Key Laboratory of Animal Models and Human Disease Mechanism of the Chinese Academy of Science & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; United Laboratory of Natural Peptide of University of Science and Technology of China, Kunming Institute of Zoology, Chinese Academy of Science, Kunming, Yunnan, 650223, China; Sino-African Joint Research Center, Chinese Academy of Science, Wuhan, Hubei, 430074, China. Electronic address: lvqm@mail.kiz.ac.cn.
authors.forename: Qiumin
authors.initials: Q
authors.lastname: Lu
mesh_terms: D000818:Animals; D004175:Diptera / Q000201:enzymology / Q000235:genetics*; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins; D012469:Salivary Glands; D012471:Salivary Proteins and Peptides / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D005343:Fibrinolytic Agents; D019476:Insect Proteins; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides
keywords: Anticoagulant; Fibrinogenolytic enzyme; Horsefly; SCP domain; Salivary gland