Leichenko E V, Monastirnaya M M, Zelepuga E A, Tkacheva E S, Isaeva M P, Likhatskaya G N, Anastyuk S D, Kozlovskaya E P
Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences, 100 Let Vladivostoku Prosp., 159, Vladivostok, 690022, Russia.
Acta Naturae. 2014 Oct;6(4):89-98.
Several new actinoporin isoforms with molecular weights of 18995.5 to 19398.7 Da exhibiting a high hemolytic activity were isolated from the tropical sea anemone Heteractis crispa using a combination of liquid chromatography techniques. The actinoporins were demonstrated to occur as mono-, di-, and trimers in aqueous solutions. The sequences of the genes encoding actinoporins were identified, and the amino acid sequences of the new polypeptides belonging to the Hct-A actinoporin family were obtained. The new acinoporins differ in their isoelectric points, the number and localization of charged amino acid residues at the functionally important N-terminal fragment of the molecule, as well as in the charge of a tetrapeptide (amino acid residues 74-77) involved in an electrostatic interaction with the cytoplasmic membrane. A recombinant actinoporin, rHct-A2, with a molecular weight of 19141 Da, pI of 9.64, and hemolytic activity of 4.0 × 104 HU/mg, was obtained. The conductivity of the ion channels formed by rHct-A2 in the BLM was demonstrated to be similar to that of the native actinoporin from H. crispa. The obtained data expand knowledge on the structural and functional relationships of actinoporins and contribute to our understanding of the functioning mechanism of these molecules, which is the basis for the development of compounds with a high biomedical potential. Currently, they are considered as models for obtaining antitumor, antibacterial, and cardiac-stimulating agents.
利用液相色谱技术相结合的方法，从热带海葵卷曲异辐海葵（Heteractis crispa）中分离出了几种分子量在18995.5至19398.7 Da之间、具有高溶血活性的新放线菌素异构体。已证明放线菌素在水溶液中以单体、二聚体和三聚体形式存在。确定了编码放线菌素的基因序列，并获得了属于Hct-A放线菌素家族的新多肽的氨基酸序列。新的放线菌素在其等电点、分子功能重要的N端片段上带电荷氨基酸残基的数量和定位以及与细胞质膜发生静电相互作用的四肽（氨基酸残基74 - 77）的电荷方面存在差异。获得了一种重组放线菌素rHct-A2，其分子量为19141 Da，pI为9.64，溶血活性为4.0×104 HU/mg。已证明rHct-A2在BLM中形成的离子通道的电导率与卷曲异辐海葵天然放线菌素的电导率相似。所获得的数据扩展了对放线菌素结构与功能关系的认识，并有助于我们理解这些分子的作用机制，这是开发具有高生物医学潜力化合物的基础。目前，它们被视为获取抗肿瘤、抗菌和心脏刺激剂的模型。
