An in vitro amino acid incorporation method for assessing the status of in vivo protein synthesis

Thaddeus Nowak, Elizabeth R. Carty, W. David Lust, Janet V. Passonneau

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A quantitative in vitro amino acid incorporation assay is described which can be used to assess the status of in vivo protein synthesis. The preparation and incubation conditions employed result in constant precursor specific activity and limit amino acid incorporation to completion of nascent peptide chains. Results obtained with this method correlate well with measurements of polyribosome profiles using sucrose gradient centrifugation. The assay is easily applied to a large number of samples, and requires only a fraction of the time and tissue necessary for conventional measures of polysome aggregation. The method has been found suitable for studies of protein synthesis in mouse brain and liver, and in gerbil brain, but not in mouse kidney. Products of in vitro protein synthesis can be separated by standard electrophoretic techniques, allowing a characterization of proteins whose mRNAs are actively translated in vivo.

Original languageEnglish (US)
Pages (from-to)285-292
Number of pages8
JournalAnalytical Biochemistry
Volume136
Issue number2
DOIs
StatePublished - Feb 1 1984
Externally publishedYes

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Amino Acids
Polyribosomes
Assays
Brain
Proteins
Centrifugation
Gerbillinae
Liver
Sucrose
Agglomeration
Tissue
Kidney
Messenger RNA
Peptides
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

An in vitro amino acid incorporation method for assessing the status of in vivo protein synthesis. / Nowak, Thaddeus; Carty, Elizabeth R.; Lust, W. David; Passonneau, Janet V.

In: Analytical Biochemistry, Vol. 136, No. 2, 01.02.1984, p. 285-292.

Research output: Contribution to journalArticle

Nowak, Thaddeus ; Carty, Elizabeth R. ; Lust, W. David ; Passonneau, Janet V. / An in vitro amino acid incorporation method for assessing the status of in vivo protein synthesis. In: Analytical Biochemistry. 1984 ; Vol. 136, No. 2. pp. 285-292.
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