Hct-A Is a New Actinoporin Family from the Heteractis Crispa Sea Anemone

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Abstract

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.

About the authors

E. V. Leichenko

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Author for correspondence.
Email: 969844@gmail.com
Россия

M. M. Monastirnaya

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

E. A. Zelepuga

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

E. S. Tkacheva

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

M. P Isaeva

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

G. N. Likhatskaya

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

S. D. Anastyuk

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

E. P. Kozlovskaya

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Россия

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Copyright (c) 2014 Leichenko E.V., Monastirnaya M.M., Zelepuga E.A., Tkacheva E.S., Isaeva M.P., Likhatskaya G.N., Anastyuk S.D., Kozlovskaya E.P.

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