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New actinoporins from sea anemone Heteractis crispa: Cloning and functional expression

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Abstract

A new actinoporin Hct-S4 (molecular mass 19,414 ± 10 Da) belonging to the sphingomyelin-inhibited α-pore forming toxin (α-PFT) family was isolated from the tropical sea anemone Heteractis crispa (also called Radianthus macrodactylus) and purified by methods of protein chemistry. The N-terminal nucleotide sequence (encoding 20 amino acid residues) of actinoporin Hct-S4 was determined. Genes encoding 18 new isoforms of H. crispa actinoporins were cloned and sequenced. These genes form a multigene Hct-S family characterized by presence of N-terminal serine in the mature proteins. Highly conserved residues comprising the aromatic phosphorylcholine-binding site and significant structure-function changes in the N-terminal segment (10–27 amino acid residues) of actinoporins were established. Two expressed recombinant actinoporins (rHct-S5 and rHct-S6) were one order less hemolytically active than native actinoporins.

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Abbreviations

a.a.:

amino acid residue

BLM:

bilayer lipid membrane

GST:

glutathione S-transferase

IPTG:

isopropyl-β-D-1-thiogalactoside

PFT:

pore forming toxin

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Authors and Affiliations

  1. Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences, pr. 100 let Vladivostoku 159, 690022, Vladivostok, Russia

    E. S. Tkacheva, E. V. Leychenko, M. M. Monastyrnaya, M. P. Issaeva, E. A. Zelepuga, S. D. Anastuk, P. S. Dmitrenok & E. P. Kozlovskaya

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  1. E. S. Tkacheva
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  2. E. V. Leychenko
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  3. M. M. Monastyrnaya
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  4. M. P. Issaeva
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  5. E. A. Zelepuga
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  6. S. D. Anastuk
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  7. P. S. Dmitrenok
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  8. E. P. Kozlovskaya
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Correspondence to M. M. Monastyrnaya.

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Original Russian Text © E. S. Tkacheva, E. V. Leychenko, M. M. Monastyrnaya, M. P. Issaeva, E. A. Zelepuga, S. D. Anastuk, P. S. Dmitrenok, E. P. Kozlovskaya, 2011, published in Biokhimiya, 2011, Vol. 76, No. 10, pp. 1387–1397.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM11-147, September 4, 2011.

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Tkacheva, E.S., Leychenko, E.V., Monastyrnaya, M.M. et al. New actinoporins from sea anemone Heteractis crispa: Cloning and functional expression. Biochemistry Moscow 76, 1131–1139 (2011). https://doi.org/10.1134/S0006297911100063

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