title: Malabarase, a serine protease with anticoagulant activity from Trimeresurus malabaricus venom.
abstract: In the present study we describe the purification and characterization of Malabarase, a serine protease from Trimeresurus malabaricus venom. Purification was achieved by gel-permeation chromatography on Sephadex G-75 followed by ion-exchange chromatography on CM Sephadex C-25. Homogeneity of Malabarase was confirmed by RP-HPLC. Malabarase is a monomer that migrated as a single protein band on SDS-PAGE under both reducing and non-reducing conditions. The molecular mass of Malabarase was determined to be 23.4 kDa using MALDI-TOF mass spectrometry. Malabarase is the first serine protease purified from T. malabaricus venom and is selective for fibrinogen. Malabarase hydrolyzes Aα and Bβ but not γ-chains of fibrinogen similar to the metalloproteases, Malabarin and Trimarin, isolated from the same venom. However, the action of Malabarase on plasma coagulation is opposite than those of Malabarin, Trimarin and the whole venom. Malabarase significantly prolonged plasma coagulation time from 152-341 s; whereas Malabarin, Trimarin, and whole venom, greatly reduce plasma clotting time from 152 to 12, 48, and 14 s, respectively. Malabarase did not show hemorrhagic or myotoxic activity. In contrast, Malabarin, Trimarin and whole venom are highly hemorrhagic and myotoxic. These observations support the specificity of Malabarase towards fibrinogen and its non-toxic nature. In conclusion, Malabarase is a fibrinogen-specific, anti-coagulant, and non-toxic serine protease. Its selective action and non-toxic nature might make it useful for treating thrombotic disorders.
authors.affiliation: Department of Studies in Bioscience, University of Mysore, Hemagangotri, Hassan-573201, Karnataka, India.
authors.forename: Raju V
authors.initials: RV
authors.lastname: Kumar
authors.forename: M
authors.initials: M
authors.lastname: Yariswamy
authors.forename: Vikram
authors.initials: V
authors.lastname: Joshi
authors.forename: K K
authors.initials: KK
authors.lastname: Dharmappa
authors.forename: Shivaprasad H
authors.initials: SH
authors.lastname: Venkatesha
authors.forename: B K
authors.initials: BK
authors.lastname: Sharath
authors.forename: Bannikuppe S
authors.initials: BS
authors.lastname: Vishwanath
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D001777:Blood Coagulation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D056148:Chromatography, Reverse-Phase; D003402:Creatine Kinase / Q000097:blood / Q000378:metabolism; D003435:Crotalid Venoms / Q000201:enzymology* / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000378:metabolism; D006470:Hemorrhage / Q000139:chemically induced; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D057057:Serine Proteases / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D012867:Skin / Q000098:blood supply / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D017841:Trimeresurus / Q000378:metabolism*; D014914:Whole Blood Coagulation Time
chemical_list: D000925:Anticoagulants; D003435:Crotalid Venoms; C029748:Trimeresurus venoms; D005340:Fibrinogen; D003402:Creatine Kinase; D057057:Serine Proteases; C581997:malabarase, Trimeresurus malabaricus