Nagaraju Shivaiah, Devaraja Sannaningaiah, Kemparaju Kempaiah
Department of Biochemistry, University of Mysore, Manasagangotri, Mysore 570 006, India.
Toxicon. 2007 Sep 1;50(3):383-93. doi: 10.1016/j.toxicon.2007.04.007. Epub 2007 Apr 24.
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.
蜘蛛毒液是蛋白质和肽类毒素的复杂混合物。透明质酸酶作为一种“扩散因子”，在蜘蛛毒液中尚未得到广泛研究。在本文中，我们描述了从帕氏希帕蛛毒液腺提取物中纯化和鉴定一种透明质酸酶的过程。透明质酸酶（HPHyal）通过在葡聚糖凝胶G - 100和CM - 葡聚糖凝胶C - 25柱上连续层析进行纯化。HPHyal已被纯化至约20倍的程度。通过基质辅助激光解吸电离飞行时间（MALDI - TOF）质谱法测得其分子量为42.26 kDa。HPHyal在37℃、pH 5.8且反应混合物中存在300 mM NaCl的条件下具有最佳活性。HPHyal对透明质酸具有绝对特异性，属于中性活性酶类。在使用针对毒液腺提取物制备的抗血清进行的蛋白质免疫印迹分析中，HPHyal显示出单一沉淀线，而毒液腺提取物显示出多条带。HPHyal间接增强了VRV - PL - VIII肌毒素的肌毒性以及出血复合物 - I的出血效力。阳离子Na⁺和K⁺增强了该酶的活性，而氯离子没有任何影响，二价阳离子则抑制酶的活性。
