Stábeli Rodrigo G, Sant'Ana Carolina D, Ribeiro Patrícia H, Costa Tássia R, Ticli Fábio K, Pires Matheus G, Nomizo Auro, Albuquerque Sérgio, Malta-Neto Natael R, Marins Mozart, Sampaio Suely V, Soares Andreimar M
Laboratório de Bioquímica do Instituto de Pesquisas em Patologias Tropicais (IPEPATRO), Universidade Federal de Rondônia, UNIR, Porto Velho-RO, Brazil.
Int J Biol Macromol. 2007 Jul 1;41(2):132-40. doi: 10.1016/j.ijbiomac.2007.01.006. Epub 2007 Jan 21.
An L-amino acid oxidase isolated from Bothrops moojeni snake venom (BmooLAAO-I) was purified to a high degree using sequential CM-Sepharose ion-exchange and phenyl-Sepharose chromatography. When analyzed by mass spectrometry, the purified BmooLAAO-I presented a molecular weight of 64,889 and 130,779 under denaturing and nondenaturing conditions, respectively. BmooLAAO-I is a homodimeric acidic glycoprotein with a pI approximately 4.7, and the N-terminal sequence shows close structural similarity to other snake venom LAAOs. This enzyme was inactivated by freezing or low pH, and secondary structural analysis by circular dichroism revealed 48% alpha-helix, 20% beta-sheet, 12% beta-turn, and 20% random coil structures. BmooLAAO-I exhibited bactericidal, antitumoral, trypanocidal, edematogenic, and platelet-aggregating activities. All of these effects were inhibited by catalase, suggesting that these biological effects are mediated by the production of H(2)O(2). BmooLAAO-I induced typical apoptotic DNA fragmentation in HL-60 cells, which was also inhibited by catalase. These results point to the potential use of BmooLAAO-I as a therapeutic agent for treatment of diseases in which induction of H(2)O(2) production can be beneficial.
从矛头蝮蛇毒液中分离出的一种L-氨基酸氧化酶（BmooLAAO-I），通过连续的CM-琼脂糖离子交换和苯基-琼脂糖层析被高度纯化。在质谱分析时，纯化后的BmooLAAO-I在变性和非变性条件下的分子量分别为64,889和130,779。BmooLAAO-I是一种同二聚体酸性糖蛋白，其pI约为4.7，N端序列与其他蛇毒LAAOs显示出紧密的结构相似性。该酶可被冷冻或低pH值灭活，圆二色性二级结构分析显示其含有48%的α-螺旋、20%的β-折叠、12%的β-转角和20%的无规卷曲结构。BmooLAAO-I表现出杀菌、抗肿瘤、杀锥虫、致水肿和血小板聚集活性。所有这些作用都被过氧化氢酶抑制，表明这些生物学效应是由H₂O₂的产生介导的。BmooLAAO-I在HL-60细胞中诱导典型的凋亡DNA片段化，这也被过氧化氢酶抑制。这些结果表明BmooLAAO-I有潜力作为一种治疗剂，用于治疗那些诱导H₂O₂产生有益的疾病。
