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

• Published in: Biochemistry (Moscow) Vol. 76; no. 10; pp. 1131 - 1139
• Main Authors: Tkacheva, E. S., Leychenko, E. V., Monastyrnaya, M. M., Issaeva, M. P., Zelepuga, E. A., Anastuk, S. D., Dmitrenok, P. S., Kozlovskaya, E. P.
• Format: Journal Article
• Language: English
• Published: Dordrecht SP MAIK Nauka/Interperiodica 01.10.2011; Springer; Springer Nature B.V
• Subjects: Amino Acid Sequence; Amino acids; Animals; Aquatic life; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Chemical plants; Cloning; Cloning, Molecular; Cnidarian Venoms - chemistry; Cnidarian Venoms - genetics; Cnidarian Venoms - isolation & purification; Cytotoxins - chemistry; Cytotoxins - genetics; Cytotoxins - isolation & purification; Gene expression; Genes; Heteractis; Invertebrates; Life Sciences; Marine; Microbiology; Molecular Sequence Data; Protection and preservation; Protein Isoforms - genetics; Proteins; Radianthus macrodactylus; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Sea Anemones - genetics; Sea Anemones - metabolism; Sequence Alignment; Structure-Activity Relationship; Toxins; recombinant actinoporin; multigene Hct-S family; actinoporin; sea anemone; hemolytic activity, structure-function analysis
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1134/S0006297911100063

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.