Peptide Mapping Study of R17 Coat Protein Synthesized in Vitro

D. J W Burns, P. L. Bergquist

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Translation of R17 ribonucleic acid (RNA) by Escherichia coli cell-free extracts was examined by studying lysine incorporation into soluble tryptic peptides from the in vitro coat protein. A sensitive thin-layer peptide mapping technique showed that several minor peptides containing lysine were present in tryptic digests both of protein coat synthesized in vitro and of natural coat protein. Radioactive double labeling of these minor peptides showed that they arose from irregularities of tryptic cleavage during protein digestion rather than from errors in translation. Five unique lysyl tryptic peptide sequences were shown to contain essentially equivalent amounts of radioactivity when the lysine content of the minor peptides was taken into account in a quantitative estimation of total lysine incorporation in vitro. This result demonstrates a high fidelity of in vitro translation of R17 RNA in the E. coli cell-free system.

LanguageEnglish
Pages1310-1317
Number of pages8
JournalBiochemistry
Volume9
Issue number6
DOIs
Publication statusPublished - 1 Mar 1970
Externally publishedYes

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Peptide Mapping
Capsid Proteins
Lysine
Peptides
Escherichia coli
RNA
Cell-Free System
Cell Extracts
Radioactivity
Proteolysis
Labeling
In Vitro Techniques
Proteins

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Burns, D. J W ; Bergquist, P. L. / Peptide Mapping Study of R17 Coat Protein Synthesized in Vitro. In: Biochemistry. 1970 ; Vol. 9, No. 6. pp. 1310-1317.
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Peptide Mapping Study of R17 Coat Protein Synthesized in Vitro. / Burns, D. J W; Bergquist, P. L.

In: Biochemistry, Vol. 9, No. 6, 01.03.1970, p. 1310-1317.

Research output: Contribution to journalArticleResearchpeer-review

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