Citation

Nagaraju S, Devaraja S, Kemparaju K. Toxicon 2007; 50(3): 383-393.

Affiliation

Department of Biochemistry, University of Mysore, Manasagangotri, Mysore 570 006, India.

Copyright

(Copyright © 2007, Elsevier Publishing)

DOI

10.1016/j.toxicon.2007.04.007

PMID

17548101

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.

Language: en