title: New Actinoporins from sea anemone Heteractis crispa: cloning and functional expression.
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.
authors.affiliation: Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok, Russia.
authors.forename: E S
authors.initials: ES
authors.lastname: Tkacheva
authors.forename: E V
authors.initials: EV
authors.lastname: Leychenko
authors.forename: M M
authors.initials: MM
authors.lastname: Monastyrnaya
authors.forename: M P
authors.initials: MP
authors.lastname: Issaeva
authors.forename: E A
authors.initials: EA
authors.lastname: Zelepuga
authors.forename: S D
authors.initials: SD
authors.lastname: Anastuk
authors.forename: P S
authors.initials: PS
authors.lastname: Dmitrenok
authors.forename: E P
authors.initials: EP
authors.lastname: Kozlovskaya
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003064:Cnidarian Venoms / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003603:Cytotoxins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012615:Sea Anemones / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
chemical_list: D003064:Cnidarian Venoms; D003603:Cytotoxins; D020033:Protein Isoforms; D011994:Recombinant Proteins