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Membrane destabilization by ricin

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Abstract

Ricin is a promising candidate for the treatment of cancer because it can be selectively targeted to tumor cells via linkage to monoclonal antibodies. Biochemical evidence suggests that escape of ricin or its ribosome-inactivating subunit from an intracellular compartment is mediated by retrograde transport to the endoplasmic reticulum and subsequent direction into the ER-associated degradation pathway. Alternatively, lipase activity of ricin may facilitate leakage from endocytic vesicles. We have observed ricin-mediated release of macromolecular dyes from lipid vesicles that mimic the composition of endosomal membranes. Release of small molecules occurs to the same extent, suggesting an all-or-none mechanism due to bilayer destabilization. The level of accompanying membrane fusion depends on vesicle composition. Since it takes 24 h of incubation before the first traces of lysolipids are detectable by matrix-assisted laser desorption/ionization mass spectrometry, membrane destabilization is not due to the lipase activity of ricin.

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Abbreviations

CF:

Carboxyfluorescein

DPhPC:

Diphytanoyl-phosphatidylcholine

DPA:

Dipicolinic acid

EDTA:

Ethylendiamine-tetracetate

ER:

Endoplasmic reticulum

ERAD:

ER-associated degradation

FRET:

Fluorescence-resonance energy transfer

GM1 :

Monosialoganglioside

MALDI-MS:

Matrix-assisted laser desorption/ionization mass spectrometry

MES:

2-Morpholino-ethanesulfonic acid

NBD-PE:

N-(7-nitro-2,1,3-benzoxadiazol-4-yl)-phosphatidylethanolamine)

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PG:

Phosphatidylglycerol

Rh-PE:

N-(lissamine rhodamine B sulfonyl)-phosphatidylethanolamine

RIP:

Ribosome-inactivating protein

RTA:

A-chain of ricin

RTB:

B-chain of ricin

TES:

N-[Tris-(hydroxymethyl)-methyl]-2-aminoethansulfonic acid

TOF:

Time-of-flight

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Acknowledgements

The work was supported by the Deutsche Forschungsgemeinschaft (Po 533/1–2; Po 533/7–1).

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Authors and Affiliations

  1. Campus Berlin Buch, Forschungsinstitut für Molekulare Pharmakologie, Robert Roessle Str. 10, 13125 , Berlin, Germany

    Jan Sun, Oxana O. Krylova, Eberhard Krause & Peter Pohl

  2. Neuroscience Research Center, Charite, Humboldt-Universität, Schumannstr. 20/21, 10117 , Berlin, Germany

    Elena E. Pohl

  3. Institute of Transplantology and Artificial Organs, Shchukinskaya St. 1, 123182 , Moscow, Russia

    Igor I. Agapov & Alexander G. Tonevitsky

  4. Lehrstuhl für Experimentelle Biophysik, Institut für Biologie, Humboldt-Universität, Invalidenstr. 43, 10115 , Berlin, Germany

    Peter Pohl

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  1. Jan Sun
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  2. Elena E. Pohl
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  3. Oxana O. Krylova
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  7. Peter Pohl
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Correspondence to Peter Pohl.

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Sun, J., Pohl, E.E., Krylova, O.O. et al. Membrane destabilization by ricin. Eur Biophys J 33, 572–579 (2004). https://doi.org/10.1007/s00249-004-0400-9

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  • DOI: https://doi.org/10.1007/s00249-004-0400-9

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