Suzuki Mieko, Itoh Takeshi, Anuruddhe B M, Bandaranayake I K, Shirani Ranasinghe J G, Athauda Seranath B P, Moriyama Akihiko
Nagoya City University, Japan.
Biomed Res. 2010 Feb;31(1):71-81. doi: 10.2220/biomedres.31.71.
To examine the molecular diversity of the venom proteins of the Russell's viper (Daboia russellii russellii) and the Indian cobra (Naja naja) in Sri Lanka, we isolated 38 venom proteins through a combination of anion exchange chromatography followed by reversed-phase high performance liquid chromatography. From the venom of D. r. russellii we isolated 15 proteins: 5 isozymes of phospholipase A(2) (PLA(2)), 4 serine proteases, 2 C-type lectin-like proteins, 2 L-amino acid oxidases, 1 cysteine-rich secretory protein (CRISP), and 1 metalloproteinase. From the venom of N. naja we isolated 23 proteins: 10 isoforms of cytotoxins (CTX), 7 PLA(2) isozymes, 2 muscarinic toxinlike proteins, 2 CRISPs, 1 nerve growth factor, and 1 new thrombin-like serine protease. Most of these proteins contained new amino acid sequences for each species, indicating molecular diversity in venom proteins. The entire amino acid sequences of PLA(2)3 from D. r. russellii and CTX7 from N. naja were determined. Additionally, the polymorphic amino acid residues of PLA(2)3 were preferentially localized on the potential antigenic sites. While 2 types of PLA(2) (N and S types) were found in D. r. russellii (India) and D. r. siamensis (Java), all the PLA(2)s from D. r. siamensis (Burma) were N type, and those from D. r. russellii (Sri Lanka) were primarily S type.
为研究斯里兰卡罗素蝰蛇（Daboia russellii russellii）和印度眼镜蛇（Naja naja）毒液蛋白的分子多样性，我们通过阴离子交换色谱结合反相高效液相色谱法分离出38种毒液蛋白。从罗素蝰蛇的毒液中，我们分离出15种蛋白：5种磷脂酶A(2)（PLA(2)）同工酶、4种丝氨酸蛋白酶、2种C型凝集素样蛋白、2种L-氨基酸氧化酶、1种富含半胱氨酸的分泌蛋白（CRISP）和1种金属蛋白酶。从印度眼镜蛇的毒液中，我们分离出23种蛋白：10种细胞毒素（CTX）同工型、7种PLA(2)同工酶、2种毒蕈碱毒素样蛋白、2种CRISP、1种神经生长因子和1种新的类凝血酶丝氨酸蛋白酶。这些蛋白中的大多数对每个物种而言都包含新的氨基酸序列，表明毒液蛋白存在分子多样性。测定了罗素蝰蛇的PLA(2)3和印度眼镜蛇的CTX7的完整氨基酸序列。此外，PLA(2)3的多态性氨基酸残基优先定位在潜在抗原位点上。在印度的罗素蝰蛇和爪哇的暹罗罗素蝰蛇中发现了2种类型的PLA(2)（N型和S型），而缅甸的暹罗罗素蝰蛇的所有PLA(2)均为N型，斯里兰卡的罗素蝰蛇的PLA(2)主要为S型。
