Immunotoxins containing A‐chain of mistletoe lectin I are more active than immunotoxins with ricin A‐chain

Tonevitsky, Alexander G.; Агапов, И. И.; Shamshiev, Abdijapar; Temyakov, D. E.; Pohl, Peter; Kirpichnikov, M. P.

doi:10.1016/0014-5793(96)00803-4

Cited by 39 publications

(13 citation statements)

References 18 publications

(25 reference statements)

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“…Mistletoe lectin and its subunits were isolated as previously described [8]. The chimeric toxin consisting of MLA and B subunit of ricin was prepared as previously described [9].…”

Section: Methodsmentioning

confidence: 99%

Mistletoe lectin dissociates into catalytic and binding subunits before translocation across the membrane to the cytoplasm

et al. 1999

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Hybridomas producing monoclonal antibodies (mAbs) against the mistletoe lectin A-chain (MLA) were obtained to investigate the intracellular routing and translocation of ribosome-inactivating proteins. Anti-MLA mAb MNA5 did not bind the holotoxin but interacted with isolated MLA. This epitope was not recognized upon MLA denaturation or conjugation of MLA with the ricin binding subunit (RTB). Furthermore, the mAbs did not appreciably react with a panel of MLA synthetic octapeptides linked to the surface of polyethylene pins. A study of the cytotoxicity of mistletoe lectin, ricin, and chimeric toxin MLA/RTB for the hybridomas revealed that interchain disulfide bond reduction and subunit dissociation are required for cytotoxic activity of mistletoe lectin.z 1999 Federation of European Biochemical Societies.

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“…Mistletoe lectin and its subunits were isolated as previously described [8]. The chimeric toxin consisting of MLA and B subunit of ricin was prepared as previously described [9].…”

Section: Methodsmentioning

confidence: 99%

Mistletoe lectin dissociates into catalytic and binding subunits before translocation across the membrane to the cytoplasm

et al. 1999

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“…ML and its subunits were isolated as previously described [5]. The chimerical toxin consisting of MLA and B-subunit of ricin was prepared as previously described [8].…”

Section: Protein Puri¢cation and Hybridoma Productionmentioning

confidence: 99%

“…Ricin and ML cytotoxicity was estimated by cell survival in the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test as earlier described [5].…”

Section: Binding and Cytotoxic Assaysmentioning

confidence: 99%

“…The interaction of monAbs with ML and subunits was analyzed in ELISA; the data were read with a spectrophotometer`Multiscan' (LabSystems, Finland). Ricin and ML cytotoxicity was estimated by cell survival in the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test as earlier described [5].…”

Section: Binding and Cytotoxic Assaysmentioning

confidence: 99%

“…MLA can closely interact with phospholipid bilayer because MLA has been shown to induce aggregation and fusion of liposomes in vitro [5,6]. Conformational changes of toxins during their intracellular tra¤cking remain unclear.…”

Section: Introductionmentioning

confidence: 99%

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Mistletoe lectin A‐chain unfolds during the intracellular transport

et al. 1999

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Protein conformation during intracellular routing and translocation of the ribosome-inactivating proteins was investigated on hybridomas producing monoclonal antibodies (monAbs) against mistletoe lectin (ML). Decrease in the toxin activity towards these hybridomas is accounted for by the intracellular interaction of monAbs and the toxin resulting in the interruption of enzymatic subunit translocation into the cytosol. Obtained monAbs interacted with denatured ML A-chain (MLA) and a panel of MLA synthetic octapeptides linked to the surface of polyethylene pins. Enzyme-linked immunosorbent assay (ELI-SA) shows that monAbs recognize five epitopes in denatured MLA. Treatment of MLA by 3 M of guanidine hydrochloride leads to appearance of the epitopes. Hybridoma TA7 has been shown to be insensitive to cytotoxic action of ML. TA7 monAb as we have shown recognizes epitope 101^105, FTGTT, and inhibits the liposome aggregation induced by MLA. A study of the cytotoxicity of ML and ricin for the hybridomas revealed that the unfolding of A-chain is probably required for intracellular transport and cytotoxic activity of ML.z 1999 Federation of European Biochemical Societies.

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