In vitro amino acid incorporation by the post-mitochondrial supernatant from rat liver

Arlan Richardson, Evelyn McGown, L. M. Henderson, Patricia B. Swan

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Interest in the use of the post-mitochondrial supernatant (PM-supernatant) for the study of in vitro amino acid incorporation arose from experiments indicating that the ribosomal aggregation of the PM-supernatant system more nearly represented that of the intact cell than did other ribosomal preparations. The PM-supernatant system was found to be 25 or 50 % more active than the microsomal and ribosomal amino acid incorporation systems. For PM-supernatant in vitro incorporation of l-[14C]valine into acid insoluble material, ATP, GTP, an energy generating system, Mg2+, K+, and a mixture of l-amino acids were necessary. Sephadex G-25 column chromatography of the PM-supernatant, pH 7.4, and presence of glutathione during homogenization of the tissue were necessary for optimal PM-supernatant in vitro incorporation. Under these conditions the rate of l-valine incorporation by the PM-supernatant from 0.17 to 0.20 nmoles/min per mg RNA for a liver from a fed rat. Fasting (20 h) prior to sacrifice, resulted in a 30-40 % decrease in amino acid incorporation by the PM-supernatant as well as a decrease in ribosomal aggregation. The decrease in PM-supernatant incorporation by fasting was found to be associated with both polysomal and cell sap factors.

Original languageEnglish (US)
Pages (from-to)468-477
Number of pages10
JournalBBA Section Nucleic Acids And Protein Synthesis
Volume254
Issue number3
DOIs
StatePublished - Dec 30 1971

ASJC Scopus subject areas

  • Medicine(all)

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