Lethal and non-lethal responses of spermatozoa from a wide variety of vertebrates and invertebrates to lysenin, a protein from the coelomic fluid of the earthwormEisenia foetida

Kobayashi, Hideshi; Sekizawa, Yoshiyuki; Aizu, Masako; Umeda, Masato

doi:10.1002/(sici)1097-010x(20000401)286:5<538::aid-jez12>3.0.co;2-w

Cited by 26 publications

(13 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As a result of pore formation, lysenin causes leakage of liposomes and lysis of erythrocytes in a sphingomyelin-dependent manner [14]. The cytolytic activity of lysenin has also been claimed to cause death of cultured mammalian cells and vertebrate spermatozoa [19,20]. It is of interest, however, that the first bioactivity of lysenin to be discovered was contraction of strips of rat aorta.…”

Section: Introductionmentioning

confidence: 99%

Lysenin-His, a sphingomyelin-recognizing toxin, requires tryptophan 20 for cation-selective channel assembly but not for membrane binding

Kwiatkowska

Hordejuk

Szymczyk

et al. 2007

Molecular Membrane Biology

View full text Add to dashboard Cite

Lysenin is 297 amino acid long toxin derived from the earthworm Eisenia foetida which specifically recognizes sphingomyelin and induces cell lysis. We synthesized lysenin gene supplemented with a polyhistidine tag, subcloned it into the pT7RS plasmid and the recombinant protein was produced in Escherichia coli. In order to obtain lysenin devoid of its lytic activity, the protein was mutated by substitution of tryptophan 20 by alanine. The recombinant mutant lysenin-His did not evoke cell lysis, although it retained the ability to specifically interact with sphingomyelin, as demonstrated by immunofluorescence microscopy and by dot blot lipid overlay and liposome binding assays. We found that the lytic activity of wild-type lysenin-His was correlated with the protein oligomerization during interaction with sphingomyelin-containing membranes and the amount of oligomers was increased with an elevation of sphingomyelin/lysenin ratio. Blue native gel electrophoresis indicated that trimers can be functional units of the protein, however, lysenin hexamers and nanomers were stabilized by chemical cross-linking of the protein and by sodium dodecyl sulfate. When incorporated into planar lipid bilayers, wild type lysenin-His formed cation-selective channels in a sphingomyelin-dependent manner. We characterized the channel activity by establishing its various open/closed states. In contrast, the mutant lysenin-His did not form channels and its correct oligomerization was strongly impaired. Based on these results we suggest that lysenin oligomerizes upon interaction with sphingomyelin in the plasma membrane, forming cation-selective channels. Their activity disturbs the ion balance of the cell, leading eventually to cell lysis.

show abstract

Section: Introductionmentioning

confidence: 99%

Lysenin-His, a sphingomyelin-recognizing toxin, requires tryptophan 20 for cation-selective channel assembly but not for membrane binding

Kwiatkowska

Hordejuk

Szymczyk

et al. 2007

Molecular Membrane Biology

View full text Add to dashboard Cite

show abstract

“…Lysenin induces hemolysis and has cytotoxicity to vertebrate spermatozoa as well as cultured mammalian cells (12,16,18). Lysenin also induces membrane damage of liposomes in a SM-dependent manner (12).…”

mentioning

confidence: 99%

Oligomerization and Pore Formation of a Sphingomyelin-specific Toxin, Lysenin

Yamaji‐Hasegawa

Makino

Baba

et al. 2003

Journal of Biological Chemistry

Self Cite

121

146

View full text Add to dashboard Cite

Lysenin is a novel protein derived from coelomic fluid of the earthworm Eisenia foetida, which specifically recognizes sphingomyelin and induces cytolysis. The mechanism underlying lysenin-induced cell lysis has not been clarified. In this report we studied the interaction of lysenin with red blood cells as well as artificial liposomes. Our results showed that lysenin bound membranes and assembled to SDS-resistant oligomers in a sphingomyelin-dependent manner, leading to the formation of pores with a hydrodynamic diameter of ϳ3 nm. Antibody scanning analysis suggested that the Cterminal region of lysenin was exposed, whereas the N-terminal was hidden in the isolated oligomer complex. Differential scanning calorimetry revealed that lysenin interacted with both hydrophilic head group and hydrophobic hydrocarbon tails of sphingomyelin. Oligomerization but not binding was affected by the amide-linked fatty acid composition of sphingomyelin, suggesting the role of membrane fluidity in the oligomerization step.

show abstract

“…Lysenin consists of 297 amino acids with a molecular weight of 41 kDa. Lysenin induces hemolysis and is toxic to vertebrate spermatozoa, amphibian larvae, and cultured mammalian cells, such as normal spleen cells, colon cancer cells, and breast cancer cells, and also amphibian larvae [10,13,14,15]. Opper et al demonstrated by flow cytometry and immunocytochemistry that the highest amount of lysenin is expressed in the cell called chloragocytes, which is one subgroup of earthworm immune cells also called coelomocyte [16].…”

Section: Structure Of Lysenin and Induction Of Hemolysismentioning

confidence: 99%

Earthworm-Derived Pore-Forming Toxin Lysenin and Screening of Its Inhibitors

Sukumwang

Umezawa

2013

Toxins

View full text Add to dashboard Cite

Lysenin is a pore-forming toxin from the coelomic fluid of earthworm Eisenia foetida. This protein specifically binds to sphingomyelin and induces erythrocyte lysis. Lysenin consists of 297 amino acids with a molecular weight of 41 kDa. We screened for cellular signal transduction inhibitors of low molecular weight from microorganisms and plants. The purpose of the screening was to study the mechanism of diseases using the obtained inhibitors and to develop new chemotherapeutic agents acting in the new mechanism. Therefore, our aim was to screen for inhibitors of Lysenin-induced hemolysis from plant extracts and microbial culture filtrates. As a result, we isolated all-E-lutein from an extract of Dalbergia latifolia leaves. All-E-lutein is likely to inhibit the process of Lysenin-membrane binding and/or oligomer formation rather than pore formation. Additionally, we isolated tyrosylproline anhydride from the culture filtrate of Streptomyces as an inhibitor of Lysenin-induced hemolysis.

show abstract

Lethal and non-lethal responses of spermatozoa from a wide variety of vertebrates and invertebrates to lysenin, a protein from the coelomic fluid of the earthwormEisenia foetida

Cited by 26 publications

References 24 publications

Lysenin-His, a sphingomyelin-recognizing toxin, requires tryptophan 20 for cation-selective channel assembly but not for membrane binding

Lysenin-His, a sphingomyelin-recognizing toxin, requires tryptophan 20 for cation-selective channel assembly but not for membrane binding

Oligomerization and Pore Formation of a Sphingomyelin-specific Toxin, Lysenin

Earthworm-Derived Pore-Forming Toxin Lysenin and Screening of Its Inhibitors

Contact Info

Product

Resources

About