Folding of an Enzyme into an Active Conformation While Bound as Peptidyl-tRNA to the Ribosome

Wieslaw Kudlicki, John Chirgwin, Gisela Kramer, Boyd Hardesty

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

Rhodanese bound to bacterial ribosomes as peptidyl-tRNA can be folded into an enzymatically active conformation by generating C-terminal extensions of the wild-type enzyme. Rhodanese was synthesized by coupled transcription/translation in a cell-free Escherichia coli system from plasmids containing the coding sequences for the wild-type enzyme or its C-terminally extended mutants. Two proteins with extensions of 23 amino acids or longer were enzymatically active while bound to the ribosomes whereas wild-type protein and a 13-amino acid extension were not. All forms of the enzyme were active after termination and release of the full-length protein from the ribosomes. All five of the bacterial chaperones were required to substantially increase the specific enzymatic activity of the extended rhodanese while the nascent protein was bound to ribosomes. The results provide direct support for the hypothesis that proteins acquire tertiary structure as they are formed in ribosomes.

Original languageEnglish (US)
Pages (from-to)14284-14287
Number of pages4
JournalBiochemistry
Volume34
Issue number44
DOIs
StatePublished - Nov 1995
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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