An acidic phospholipase A<sub>2sub> with antibacterial activity from Porthidium nasutum snake venom
- 作者：Leidy Johana Vargas<sup>asup><sup> ; sup><sup>johana2104@gmail.comsup> ; Mó ; nica Londoñ ; o<sup>asup> ; Juan Carlos Quintana<sup>asup> ; Carolina Rua<sup>bsup> ; Cesar Segura<sup>csup> ; Bruno Lomonte<sup>dsup> ; Vitelbina Nú ; ñ ; ez<sup>asup><sup> ; sup><sup>esup>
- 关键词：PLA2 ; Phospholipase A2 ; PnPLA2 ; Phospholipase A2 of Porthidium nasutum ; MIC ; Minimum inhibitory concentration ; MBC ; Minimal bactericidal concentration
- 刊名：Comparative Biochemistry and Physiology ; Part B
- 出版年：2012
- 期刊代码：50_10964959
- 类别：bio
- 出版时间：April, 2012
- 卷：161
- 期：4
- 页码：341-347
- 文件大小：894 K
摘要
Snake venoms are complex mixtures of proteins among which both basic and acidic phospholipases A<sub>2sub> (PLA<sub>2sub>s) can be found. Basic PLA<sub>2sub>s are usually responsible for major toxic effects induced by snake venoms, while acidic PLA<sub>2sub>s tend to have a lower toxicity. A novel PLA<sub>2sub>, here named PnPLA<sub>2sub>, was purified from the venom of Porthidium nasutum by means of RP-HPLC on a C18 column. PnPLA<sub>2sub> is an acidic protein with a pI of 4.6, which migrates as a single band under both non-reducing and reducing conditions in SDS-PAGE. PnPLA<sub>2sub> had a molecular mass of 15,802.6 Da, determined by ESI-MS. Three tryptic peptides of this protein were characterized by HPLC-nESI-MS/MS, and N-terminal sequencing by direct Edman degradation showing homology to other acidic PLA<sub>2sub>s from viperid venoms. PnPLA<sub>2sub> displayed indirect hemolytic activity in agarose erythrocyte-egg yolk gels and bactericidal activity against Staphylococcus aureus in a dose-dependent manner, with a MIC and MBC of 32 渭g/mL. In addition, PnPLA<sub>2sub> showed a potent inhibitory effect on platelet aggregation with doses up to 40 渭g/mL. This acidic PLA<sub>2sub>, in contrast to basic enzymes isolated from other viperid snake venoms, was not cytotoxic to murine skeletal muscle myoblasts C<sub>2sub>C<sub>12sub>. This is the first report on a bactericidal protein of Porthidium nasutum venom.