Isolation, properties and partial amino acid sequence of a new actinoporin from the sea anemone Radianthus macrodactylus

Elena V. Klyshko; Marina P. Issaeva; Margarita M. Monastyrnaya; Anna P. Il'yna; Konstantin V. Guzev; Tatyana I. Vakorina; Pavel S. Dmitrenok; Tatyana A. Zykova; Emma P. Kozlovskaya

doi:10.1016/j.toxicon.2004.06.006

Isolation, properties and partial amino acid sequence of a new actinoporin from the sea anemone Radianthus macrodactylus

Elena V. Klyshko
, Marina P. Issaeva
, Margarita M. Monastyrnaya
, Anna P. Il'yna
, Konstantin V. Guzev
, Tatyana I. Vakorina
, Pavel S. Dmitrenok
, Tatyana A. Zykova
, Emma P. Kozlovskaya

Research output: Contribution to journal › Article › peer-review

35 Scopus citations

Abstract

A new cytolytic toxin, actinoporin RTX-S II, was isolated from the sea anemone Radianthus macrodactylus with a high degree of purity by a combination of gel filtration, ion-exchange and reverse-phase chromatography. RTX-S II has molecular mass of 19,280Da and isoelectric point of 10.0. The hemolytic activity of RTX-S II is inhibited by sphingomyelin. RTX-S II had an LD₅₀ of 70 mg/kg, and is lacking in phospholipase activity. The amino acid composition of this protein contains a high amount of basic and non-polar amino acids and no cysteine. The N-terminal sequence of RTX-S II was determined. The partial amino acid sequence (141 aa) of RTX-S II was deduced based on the cDNA sequence obtained with two oligonucleotides encoding the N-terminal portion of RTX-S II and the internal conserved cytolysin peptide by PCR. A comparison of the RTX-S II cDNA sequence and the rtx-s II gene obtained with the same PCR primers indicates that they are 100% identical at the nucleotide level. It shows that no introns are present in the corresponding region of the rtx-s II gene. Multiple alignments of RTX-S II with known sequences of actinoporins show that RTX-S II is highly homologous to magnificalysin II from Heteractis magnifica. The predicted secondary structure of RTX-S II is predominantly anti-parallel β-structure, which is in good agreement with experimental data obtained from other sea anemones-actinoporins.

Original language	English (US)
Pages (from-to)	315-324
Number of pages	10
Journal	Toxicon
Volume	44
Issue number	3
DOIs	https://doi.org/10.1016/j.toxicon.2004.06.006
State	Published - Sep 1 2004
Externally published	Yes

Bibliographical note

Funding Information:
The authors thank Dr I. Nazimov (Institute of Bioorganic Chemistry, the Russian Academy of Sciences, Moscow) for automated N-terminal amino acid sequence analyses of RTX-S II, and Mr A. Shvedov who translated this paper into English. This work was supported by the RFBR grant no. 02-04-49486, the program of Presidium of RAS “Molecular and cell biology” grant no. 03-1-0-05-002 and FEBRAS grant.

Keywords

Actinoporins
Cytolysins
Hemolytic activity
Primary and secondary structures
Sea anemones
Sphingomyelin

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10.1016/j.toxicon.2004.06.006

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title = "Isolation, properties and partial amino acid sequence of a new actinoporin from the sea anemone Radianthus macrodactylus",

abstract = "A new cytolytic toxin, actinoporin RTX-S II, was isolated from the sea anemone Radianthus macrodactylus with a high degree of purity by a combination of gel filtration, ion-exchange and reverse-phase chromatography. RTX-S II has molecular mass of 19,280Da and isoelectric point of 10.0. The hemolytic activity of RTX-S II is inhibited by sphingomyelin. RTX-S II had an LD50 of 70 mg/kg, and is lacking in phospholipase activity. The amino acid composition of this protein contains a high amount of basic and non-polar amino acids and no cysteine. The N-terminal sequence of RTX-S II was determined. The partial amino acid sequence (141 aa) of RTX-S II was deduced based on the cDNA sequence obtained with two oligonucleotides encoding the N-terminal portion of RTX-S II and the internal conserved cytolysin peptide by PCR. A comparison of the RTX-S II cDNA sequence and the rtx-s II gene obtained with the same PCR primers indicates that they are 100\% identical at the nucleotide level. It shows that no introns are present in the corresponding region of the rtx-s II gene. Multiple alignments of RTX-S II with known sequences of actinoporins show that RTX-S II is highly homologous to magnificalysin II from Heteractis magnifica. The predicted secondary structure of RTX-S II is predominantly anti-parallel β-structure, which is in good agreement with experimental data obtained from other sea anemones-actinoporins.",

keywords = "Actinoporins, Cytolysins, Hemolytic activity, Primary and secondary structures, Sea anemones, Sphingomyelin",

author = "Klyshko, \{Elena V.\} and Issaeva, \{Marina P.\} and Monastyrnaya, \{Margarita M.\} and Il'yna, \{Anna P.\} and Guzev, \{Konstantin V.\} and Vakorina, \{Tatyana I.\} and Dmitrenok, \{Pavel S.\} and Zykova, \{Tatyana A.\} and Kozlovskaya, \{Emma P.\}",

note = "Funding Information: The authors thank Dr I. Nazimov (Institute of Bioorganic Chemistry, the Russian Academy of Sciences, Moscow) for automated N-terminal amino acid sequence analyses of RTX-S II, and Mr A. Shvedov who translated this paper into English. This work was supported by the RFBR grant no. 02-04-49486, the program of Presidium of RAS {\textquotedblleft}Molecular and cell biology{\textquotedblright} grant no. 03-1-0-05-002 and FEBRAS grant.",

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doi = "10.1016/j.toxicon.2004.06.006",

language = "English (US)",

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T1 - Isolation, properties and partial amino acid sequence of a new actinoporin from the sea anemone Radianthus macrodactylus

AU - Klyshko, Elena V.

AU - Issaeva, Marina P.

AU - Monastyrnaya, Margarita M.

AU - Il'yna, Anna P.

AU - Guzev, Konstantin V.

AU - Vakorina, Tatyana I.

AU - Dmitrenok, Pavel S.

AU - Zykova, Tatyana A.

AU - Kozlovskaya, Emma P.

N1 - Funding Information: The authors thank Dr I. Nazimov (Institute of Bioorganic Chemistry, the Russian Academy of Sciences, Moscow) for automated N-terminal amino acid sequence analyses of RTX-S II, and Mr A. Shvedov who translated this paper into English. This work was supported by the RFBR grant no. 02-04-49486, the program of Presidium of RAS “Molecular and cell biology” grant no. 03-1-0-05-002 and FEBRAS grant.

PY - 2004/9/1

Y1 - 2004/9/1

N2 - A new cytolytic toxin, actinoporin RTX-S II, was isolated from the sea anemone Radianthus macrodactylus with a high degree of purity by a combination of gel filtration, ion-exchange and reverse-phase chromatography. RTX-S II has molecular mass of 19,280Da and isoelectric point of 10.0. The hemolytic activity of RTX-S II is inhibited by sphingomyelin. RTX-S II had an LD50 of 70 mg/kg, and is lacking in phospholipase activity. The amino acid composition of this protein contains a high amount of basic and non-polar amino acids and no cysteine. The N-terminal sequence of RTX-S II was determined. The partial amino acid sequence (141 aa) of RTX-S II was deduced based on the cDNA sequence obtained with two oligonucleotides encoding the N-terminal portion of RTX-S II and the internal conserved cytolysin peptide by PCR. A comparison of the RTX-S II cDNA sequence and the rtx-s II gene obtained with the same PCR primers indicates that they are 100% identical at the nucleotide level. It shows that no introns are present in the corresponding region of the rtx-s II gene. Multiple alignments of RTX-S II with known sequences of actinoporins show that RTX-S II is highly homologous to magnificalysin II from Heteractis magnifica. The predicted secondary structure of RTX-S II is predominantly anti-parallel β-structure, which is in good agreement with experimental data obtained from other sea anemones-actinoporins.

AB - A new cytolytic toxin, actinoporin RTX-S II, was isolated from the sea anemone Radianthus macrodactylus with a high degree of purity by a combination of gel filtration, ion-exchange and reverse-phase chromatography. RTX-S II has molecular mass of 19,280Da and isoelectric point of 10.0. The hemolytic activity of RTX-S II is inhibited by sphingomyelin. RTX-S II had an LD50 of 70 mg/kg, and is lacking in phospholipase activity. The amino acid composition of this protein contains a high amount of basic and non-polar amino acids and no cysteine. The N-terminal sequence of RTX-S II was determined. The partial amino acid sequence (141 aa) of RTX-S II was deduced based on the cDNA sequence obtained with two oligonucleotides encoding the N-terminal portion of RTX-S II and the internal conserved cytolysin peptide by PCR. A comparison of the RTX-S II cDNA sequence and the rtx-s II gene obtained with the same PCR primers indicates that they are 100% identical at the nucleotide level. It shows that no introns are present in the corresponding region of the rtx-s II gene. Multiple alignments of RTX-S II with known sequences of actinoporins show that RTX-S II is highly homologous to magnificalysin II from Heteractis magnifica. The predicted secondary structure of RTX-S II is predominantly anti-parallel β-structure, which is in good agreement with experimental data obtained from other sea anemones-actinoporins.

KW - Actinoporins

KW - Cytolysins

KW - Hemolytic activity

KW - Primary and secondary structures

KW - Sea anemones

KW - Sphingomyelin

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DO - 10.1016/j.toxicon.2004.06.006

M3 - Article

C2 - 15302538

AN - SCOPUS:4043125369

SN - 0041-0101

VL - 44

SP - 315

EP - 324

JO - Toxicon

JF - Toxicon

IS - 3

ER -

Isolation, properties and partial amino acid sequence of a new actinoporin from the sea anemone Radianthus macrodactylus

Abstract

Bibliographical note

Keywords

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