title: The Hag-protease-II is a fibrin(ogen)ase from Hippasa agelenoides spider venom gland extract: purification, characterization and its role in hemostasis.
abstract: The current study describes the biochemical, biophysical and pharmacological properties of Hag-protease-II from Hippasa agelenoides spider venom gland extract. The Hag-protease-II was purified to homogeneity using gel filtration and ion-exchange chromatography. The molecular mass was found to be 28.749 kDa by MALDI-TOF mass spectrometry. PMSF abolished the activity while EDTA, EGTA, IAA and 1, 10-phenanthrolene did not. Hag-protease-II hydrolyzed casein, fibrinogen and fibrin, however it did not hydrolyze gelatin, fibronectin and collagen types- I and IV. It was non-lethal and devoid of hemorrhagic, myotoxic and edema forming activities. It dose dependently reduced re-calcification time of citrated human plasma. Strikingly; the Hag-protease-II coagulated the factor X deficient congenital human plasma. It hydrolyzed Bβ-chain but, did not degrade Aα- and γ-chains of fibrinogen while, it hydrolyzed α-polymer and α-chain but not the β-chain and γ-γ dimers of partially cross-linked fibrin clot. The Hag-protease-II induced aggregation of human platelets in PRP dose dependently, however it did not interfere in collagen induced aggregation of PRP and washed human platelets.
authors.affiliation: Department of Biochemistry, Manasagangotri, University of Mysore, Mysore 570 006, India.
authors.forename: S
authors.initials: S
authors.lastname: Devaraja
authors.forename: K S
authors.initials: KS
authors.lastname: Girish
authors.forename: Y N J
authors.initials: YN
authors.lastname: Gowtham
authors.forename: K
authors.initials: K
authors.lastname: Kemparaju
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D005260:Female; D005340:Fibrinogen / Q000737:chemistry; D006487:Hemostasis / Q000187:drug effects*; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013111:Spider Venoms / Q000201:enzymology*; D013112:Spiders / Q000201:enzymology*
chemical_list: D013111:Spider Venoms; D005340:Fibrinogen; D012697:Serine Endopeptidases