title: Purification and properties of hyaluronidase from Hippasa partita (funnel web spider) venom gland extract.
abstract: Spider venom is a complex mixture of protein and peptide toxins. Hyaluronidase a 'spreading factor' has not been studied extensively in spider venom. In this paper, we describe the purification and characterization of a hyaluronidase from Hippasa partita venom gland extract. Hyaluronidase (HPHyal) has been purified by the successive chromatography on a Sephadex G-100 and on CM-Sephadex C-25 columns. HPHyal has been purified to an extent of about approximately 20-folds. The molecular mass was found to be 42.26 kDa by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. HPHyal was optimally active at pH 5.8 at 37 degrees C and in the presence of 300 mM NaCl in the reaction mixture. HPHyal showed absolute specificity for hyaluronan and belongs to neutral active group of enzymes. HPHyal revealed single-precipitin line, while venom gland extract revealed multiple bands in Western blotting with the antiserum prepared against venom gland extract. HPHyal indirectly potentiates the myotoxicity of VRV-PL-VIII myotoxin and also the hemorrhagic potency of hemorrhagic complex-I. Cations, Na(+) and K(+) enhanced the activity and chloride ions do not have any effect while, divalent cations, inhibited the enzyme activity.
authors.affiliation: Department of Biochemistry, University of Mysore, Manasagangotri, Mysore 570 006, India.
authors.forename: Shivaiah
authors.initials: S
authors.lastname: Nagaraju
authors.forename: Sannaningaiah
authors.initials: S
authors.lastname: Devaraja
authors.forename: Kempaiah
authors.initials: K
authors.lastname: Kemparaju
mesh_terms: D000818:Animals; D004487:Edema / Q000139:chemically induced; D005260:Female; D006821:Hyaluronoglucosaminidase / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D011817:Rabbits; D013111:Spider Venoms / Q000737:chemistry / Q000201:enzymology*; D013112:Spiders / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D013111:Spider Venoms; D006821:Hyaluronoglucosaminidase