Ricin A Chain Can Be Chemically Cross-linked to the Mammalian Ribosomal Proteins L9 and L10e (∗)

Indirect immunofluorescence studies revealed that when fixed, permeabilized cultured human cells were incubated with ricin A chain, the toxin molecule localized in a staining pattern indicative of binding to the endoplasmic reticulum and to nucleoli. Chemical cross-linking experiments were performed...

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Bibliographic Details
Published inThe Journal of biological chemistry Vol. 270; no. 21; pp. 12933 - 12940
Main Authors Vater, Carol A., Bartle, Laura M., Leszyk, John D., Lambert, John M., Goldmacher, Victor S.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 26.05.1995
American Society for Biochemistry and Molecular Biology
Subjects
Amino Acid Sequence
Cell Compartmentation
Cell Nucleolus - chemistry
Cross-Linking Reagents
Endoplasmic Reticulum - chemistry
Fluorescent Antibody Technique
Humans
Molecular Sequence Data
Phosphoproteins - chemistry
Phosphoproteins - metabolism
Protein Binding
Ribosomal Protein L10
Ribosomal Proteins - chemistry
Ribosomal Proteins - metabolism
Ribosomes - chemistry
Ribosomes - metabolism
Ricin - metabolism
Sequence Analysis
Tumor Cells, Cultured
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Summary:Indirect immunofluorescence studies revealed that when fixed, permeabilized cultured human cells were incubated with ricin A chain, the toxin molecule localized in a staining pattern indicative of binding to the endoplasmic reticulum and to nucleoli. Chemical cross-linking experiments were performed to identify the cellular components that mediated the binding of ricin A chain. Conjugates were formed between 125I-labeled ricin A chain and two proteins present in preparations of total cell membranes and in samples of purified mammalian ribosomes. Specificity of the ricin A chain-ribosome interaction was demonstrated by inhibition of formation of the complexes by excess unlabeled ricin A chain, but not by excess unlabeled gelonin, another ribosome-inactivating protein. Complexes of ricin A chain cross-linked to the ribosomal proteins were purified and subjected to proteolytic digestion with trypsin. Amino acid sequencing of internal tryptic peptides enabled identification of the ricin A chain-binding proteins as L9 and L10e of the mammalian large ribosomal subunit.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.270.21.12933