Molecular characterization of L-amino acid oxidase from king cobra venom
Molecular characterization of L-amino acid oxidase from king cobra venom
复制标题
眼镜王蛇毒液 L-氨基酸氧化酶的分子表征
DOI：
10.1016/j.toxicon.2007.04.013
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发表时间：
2007-09-15
期刊：
影响因子：
2.8
通讯作者：
Zhang, Yun
中科院分区：
文献类型：
--
作者：
Jin, Yang;Lee, Wen-Hui;Zhang, Yun
通过连续凝胶过滤、离子交换和肝素层析纯化来自蛇蛇毒液的 L-氨基酸氧化酶 (Oh-LAAO)。 Oh-LAAO不诱导血小板聚集；然而，它对ADP和U46619诱导的血小板聚集有有效的抑制活性，但对凝血酶、mucetin、ristocetin和stejnulxin诱导的血小板聚集没有作用。通过RT-PCR和5'-RACE方法，从毒腺总RNA制剂中克隆出完整的Oh-LAAO cDNA。该cDNA序列含有1476bp的开放阅读框(ORF)，编码491个氨基酸的蛋白质，其中包含25个氨基酸的信号肽和466个残基的成熟蛋白质。 Oh-LAAO 的预测蛋白质序列通过 N 端和胰蛋白酶消化的内部肽测序以及肽质量指纹分析得到证实。本研究克隆并报告了编码金环蛇和多环金环蛇 LAAO 的 ORF 的 cDNA。此外，还报道了Naja atra LAAO 的部分cDNA 编码。 Oh-LAAO 与其他已知蛇毒 LAAO 具有大约 50% 的蛋白质序列同一性。系统发育分析表明 Oh-LAAO 与其他蛇毒 LAAO 的进化距离较远。 (C) 2007 Elsevier Ltd. 保留所有权利。
An L-amino acid oxidase from Ophiophagus hannah snake venom (Oh-LAAO) was purified by successive gel filtration, ion-exchange and heparin chromatography. Oh-LAAO did not induce platelet aggregation; however, it had potent inhibitory activity on platelet aggregation induced by ADP and U46619, but showed no effect on platelet aggregation induced by thrombin, mucetin, ristocetin and stejnulxin. By RT-PCR and 5'-RACE methods, the complete Oh-LAAO cDNA was cloned from the venom gland total RNA preparations. The cDNA sequence contains an open-reading frame (ORF) of 1476-bp, which encodes a protein of 491 amino acids comprising a signal peptide of 25 amino acids and 466-residue mature protein. The predicted protein sequence of Oh-LAAO was confirmed by N-terminal and trypsin-digested internal peptides sequencing together with peptide mass fingerprinting. cDNAs encoding for ORF of LAAOs from Bungarus fasciatus and B. multicinctus were cloned and reported in this study. In addition, partial cDNA encoding for Naja atra LAAO was also reported. Oh-LAAO shared approximately 50% protein sequence identity with other known snake venom LAAOs. Phylogenetic analysis indicated that Oh-LAAO is evolutionary distant to other snake venom LAAOs. (C) 2007 Elsevier Ltd. All rights reserved.