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- W2070569843 abstract "The crude soluble fraction of rat liver cytoplasm promotes the binding of acetylphenylalanyl-tRNA but not of Met-tRNAf to 40S subunits derived from 80S ribosomes. A protein has been extensively purified from the soluble fraction that catalyzes the template-dependent, GTP-independent binding of Met-tRNAf, acetylphenylalanyl-tRNA and phenylalanyl-tRNA but not Met-tRNAm. Purification involves fractionation with ammonium sulfate and chromatography on calcium phosphate gel, DEAE-Sephadex, carboxymethyl cellulose and Sephadex G-200. The optimum Mg2+ concentration for the binding reaction with Met-tRNAf is between 6 and 8 mm and the optimum temperature is between 10 and 15 °C. The complex formed as a result of the interaction between 40S subunits, acetylphenylalanyl-tRNA and poly(U) is functional; acetylpolyphenylalanine is synthesized when the isolated 40S-poly(U)·acetylphenylalanyl-tRNA complex is incubated with 60S subunits, phenylalanyl-tRNA, elongation factors and GTP. The crude cytoplasmic fraction, which does not stimulate the binding of Met-tRNAf, inhibits the purified factor-promoted binding of this substrate; the factor-independent, high magnesium ion-stimulated binding of Met-tRNAf to 40S subunits is also inhibited. The inhibitory activity can be resolved from the binding factor and is extensively purified by chromatography on calcium phosphate gel and carboxymethyl Sephadex and by electrofocusing. In the presence of 40S subunits, crude and purified preparations of the inhibitory activity hydrolyze Met-tRNAf but not Met-tRNAm or acetylphenylalanyl-tRNA. Free Met-tRNAf is not hydrolyzed. Incubation of hydrolase-containing preparations with the preformed 40S-·Met-tRNAf complex results in the rapid and extensive breakdown of the complex with release of acid-insoluble methionine; the formation of an 80S·substrate complex, by the addition of 60S subunits, protects particle-bound Met-tRNAf." @default.
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- W2070569843 date "1976-02-01" @default.
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- W2070569843 title "Partial purification and characterization of a binding factor specific for initiator tRNA and ribosomal 40S subunits and of an aminoacyl-tRNA hydrolase specific for 40S-bound Met-tRNAf from rat liver" @default.
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- W2070569843 doi "https://doi.org/10.1016/0003-9861(76)90090-4" @default.
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