Cytotoxicity acquired by ribosome‐inactivating proteins carried by reconstituted Sendai virus envelopes

Sargiacomo, Massimo; Barbieri, Luigi; Stirpe, Fiorenzo; Tomasi, M.

doi:10.1016/0014-5793(83)81135-1

Cited by 20 publications

(7 citation statements)

References 16 publications

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“…EF-1 and EF-2 were obtained from rat liver 'pH 5 supernatant' and resolved as described by Montanaro et al (1973). Protein synthesis assays were performed using the following methods: that of Hosokawa et al (1966) for poly(U)-directed polyphenylalanine synthesis by E. coli ribosomes; that of Sargiacomo et al (1983) for endogenous mRNA-directed protein synthesis by the rabbit reticulocyte lysate; and that of Barbieri et al (1980) for poly(U)-directed synthesis of polyphenylalanine by A. salina ribosomes. Hot-acid-insoluble radioactivity was collected as described by Montanaro et al (1978).…”

Section: Methodsmentioning

confidence: 99%

Effect of the antibiotic purpuromycin on cell-free protein-synthesizing systems

Rambelli

Brigotti

Zamboni

et al. 1989

Biochemical Journal

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Purpuromycin, an antibiotic isolated from the culture broth of Actinoplanes ianthinogenes, which is very active against Gram-positive bacteria and fungi, inhibits protein synthesis in both prokaryotic and eukaryotic cell-free systems. The ID50 was 9 microM with the endogenous mRNA-directed rabbit reticulocyte lysate, 17 microM with a poly(U)-directed system from Escherichia coli and 69 microM with a poly(U)-directed system from Artemia salina cysts. Of the three steps of elongation, purpuromycin does not affect the peptidyl-transferase reaction, inhibits the elongation factor 1 (EF-1) dependent binding of phenylalanyl-tRNA and stimulates the GTP-dependent binding of EF-2. When protein synthesis is stopped by the addition of purpuromycin, the nascent peptide chains are found in the puromycin-reactive P site. The results suggest that the mechanism of action of purpuromycin is similar to that of fusidic acid. Both antibiotics would seem to produce a stable guanine nucleotide-ribosome-EF-2 complex which allows one round of translocation but prevents, because of a common or overlapping ribosomal binding site for the two elongation factors, the subsequent EF-1-dependent binding of aminoacyl-tRNA.

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Section: Methodsmentioning

confidence: 99%

Effect of the antibiotic purpuromycin on cell-free protein-synthesizing systems

Rambelli

Brigotti

Zamboni

et al. 1989

Biochemical Journal

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“…This was confirmed by the marked cytotoxic effects which are observed when the entry into the cytoplasm was enhanced. This was obtained when RIPS were conjugated with molecules capable of binding to cell membranes (concanavalin A [lo], neoglycoproteins [50], mannose 6-phosphate [51], antibodies, see below), or included into structures such as liposomes [52], reconstituted Sendai virus envelopes [53] or erythrocyte ghosts [54] which could be fused with cells (table 3).…”

Section: Intact Cellsmentioning

confidence: 99%

Ribosome‐inactivating proteins up to date

1986

Self Cite

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Ribosome-inactivating proteins (RIPS) from plants inactivate eukaryotic ribosomes, as far as studied by rendering their 60 S subunit unable to bind elongation factor 2. These proteins seem widely distributed and possibly ubiquitous in plants. They are either type 1, those consisting of a single polypeptide chain, or type 2 (ricin and related toxins), those consisting of two chains, one of which is a galactose-binding lectin. The literature on RIPS from 1982 has been reviewed with respect to (i) the chemical and biological properties of RIPS, (ii) their use for the preparation of immunotoxins and (iii) new perspectives. Ribosome-inactivating protein

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“…Deglycosylation resulted in a decrease in M, of the Al-chain component by 600 and the AZ-chain component by 2000 as assessed by polyacrylamide gel electrophoresis [6]. All A-chain preparations produced comparable inhibition of protein synthesis in the reticulocyte lysate assay [9]. Deglycosylation of the B-chain was carried out by treatment at room temperature of isolated B-chain (2 mg/ml) in 0.1 M citrate buffer, pH 5.5, containing 0.2 M Dgalactose, 0.25% 2-mercaptoethanol and 1 mM PMSF (Sigma), either with only a-mannosidase (0.5 U/ml) for 3 days, or with endoglycosidase H (Miles Labs, 10 mu/ml) for 2 days, followed by addition of cu-mannosidase (0.18 U/ml) for a further 24 h. After deglycosylation 2 mg B-chain was filtered at a flow rate of 0.5 ml/min through 1.5 ml of a Con A-Sepharose column equilibrated with PBS containing 0.1 M galactose to remove undegraded B-chain.…”

Section: Deglycosylation Of A-and B-chainsmentioning

confidence: 96%

Selective uptake of ricin A‐chain by hepatic non‐parenchymal cells in vitro

Skilleter

Foxwell

1986

FEBS Letters

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Free ricin A-chain was actively taken up in vitro by rat liver non-parenchymal cells but not by parenchymal cells. A-chain uptake by non-parenchymal cells could be selectively inhibited by D-mannose, L-fucose or ovalbumin and was markedly decreased after partial removal of mannose residues from the oligosaccharides present in the glycoprotein by enzymic deglycosylation. Uptake of free ricin B-chain by non-parenchymal cells was greater than that by parenchymal cells but in both cases was little influenced by enzymic deglycosylation of the glycoprotein. The results are consistent with mannose receptor recognition of ricin A-chain by non-parenchymal cells and have important implications for the clinical use in vivo of antibody-ricin A-chain conjugates in cancer therapy. Ricin Deglycosylation Mannose oligosaccharide (Liver cell)

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Cytotoxicity acquired by ribosome‐inactivating proteins carried by reconstituted Sendai virus envelopes

Cited by 20 publications

References 16 publications

Effect of the antibiotic purpuromycin on cell-free protein-synthesizing systems

Effect of the antibiotic purpuromycin on cell-free protein-synthesizing systems

Ribosome‐inactivating proteins up to date

Selective uptake of ricin A‐chain by hepatic non‐parenchymal cells in vitro

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