Abstract
Three inhibitors of animal cell-free protein synthesis were isolated from barley seeds by ammonium sulfate fractionation, CM-cellulose chromatography and gel filtration on Bio-Gel P-60. The three inhibitors were all single chain, basic proteins with molecular weights about 31,000 and with very similar amino acid compositions. In the rabbit reticulocyte lysate system, they inhibited in vitro translation to 50% at concentrations between 15 and 25 ng·ml−1, while in the wheat germ system, their effect was very weak. The three translation inhibitors showed no detectable ribonuclease activity neither on ribosomal RNA from rabbit liver nor on homopolynucleotides.
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Abbreviations
- DTT:
-
dithiothreitol
- EDTA:
-
ethylenediamine tetraacetic acid, disodium salt
- FPLC:
-
fast protein liquid chromatography
- Hepes:
-
N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid
- PAP:
-
pokeweed antiviral protein
- PTH:
-
phenylthiohydantoin
- SDS:
-
sodium dodecylsulfate
- Tris:
-
2-amino-2-hydroxymethylpropane-1,3-diol
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Accepted by:H. Klenow, E. Lund andS.O. Andersen
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Asano, K., Svensson, B. & Poulsen, F.M. Isolation and characterization of inhibitors of animal cell-free protein synthesis from barley seeds. Carlsberg Res. Commun. 49, 619–626 (1984). https://doi.org/10.1007/BF02907494
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DOI: https://doi.org/10.1007/BF02907494