Holzer M, Mackessy S P
Department of Biological Sciences, University of Northern Colorado, Greeley 80639, USA.
Toxicon. 1996 Oct;34(10):1149-55. doi: 10.1016/0041-0101(96)00057-8.
Phospholipase A2 (PLA2), an enzyme found in most snake venoms, catalyzes the hydrolysis of phospholipids in biological membranes, and some have presynaptic neurotoxic activity. A synthetic substrate, 4-nitro-3-(octanoyloxy)benzoic acid, was synthesized and purified on a silica gel column using a published method. This substrate was used to develop an endpoint assay which is rapid and requires a minimum of equipment. This aqueous assay system allowed enzyme activity to be examined without the use of radioactive substrates or organic solvents, minimizing waste disposal concerns. Whole venoms, partially purified enzyme isolated from Crotalus mitchelli pyrrhus venom, tissue extracts and commercial preparations were employed as sources of PLA2. Results show that this method is a convenient and specific assay for PLA2 from several sources and is particularly suited for assaying large numbers of fractions generated during purification procedures.
磷脂酶A2（PLA2）是一种存在于大多数蛇毒中的酶，它催化生物膜中磷脂的水解，有些还具有突触前神经毒性活性。一种合成底物4-硝基-3-（辛酰氧基）苯甲酸，采用已发表的方法在硅胶柱上合成并纯化。该底物用于开发一种终点测定法，该方法快速且所需设备最少。这种水性测定系统无需使用放射性底物或有机溶剂就能检测酶活性，从而将废物处理问题降至最低。全毒液、从米切尔响尾蛇毒液中分离出的部分纯化酶、组织提取物和商业制剂用作PLA2的来源。结果表明，该方法是一种方便且特异的测定多种来源PLA2的方法，特别适合于测定纯化过程中产生的大量组分。
