The folding and unfolding behavior of ribonuclease H on the ribosome.

The Journal of Biological Chemistry
M. K. JensenSusan Marqusee

Abstract

The health of a cell depends on accurate translation and proper protein folding, whereas misfolding can lead to aggregation and disease. The first opportunity for a protein to fold occurs during translation, when the ribosome and surrounding environment can affect the nascent chain energy landscape. However, quantifying these environmental effects is challenging because ribosomal proteins and rRNA preclude most spectroscopic measurements of protein energetics. Here, we have applied two gel-based approaches, pulse proteolysis and force-profile analysis, to probe the folding and unfolding pathways of RNase H (RNH) nascent chains stalled on the prokaryotic ribosome in vitro We found that ribosome-stalled RNH has an increased unfolding rate compared with free RNH. Because protein stability is related to the ratio of the unfolding and folding rates, this increase completely accounts for the observed change in protein stability and indicates that the folding rate is unchanged. Using arrest peptide-based force-profile analysis, we assayed the force generated during the folding of RNH on the ribosome. Surprisingly, we found that population of the RNH folding intermediate is required to generate sufficient force to release a stall induc...Continue Reading

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Citations

Aug 14, 2020·Protein Science : a Publication of the Protein Society·Rageia ElfageihRenuka Kudva
Aug 21, 2020·The Journal of Biological Chemistry·Jonathan P Schlebach
May 15, 2021·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Kevin MaciubaChristian M Kaiser
May 15, 2021·Journal of Molecular Biology·Hena SandhuGunnar von Heijne
Oct 16, 2021·Nature Chemistry·Anaïs M E CassaignauJohn Christodoulou

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Methods Mentioned

BETA
protein folding
NMR
FRET

Software Mentioned

MatLab
ImageJ
SecM

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The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.

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