Cargando…
Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy
The folding of many proteins can begin during biosynthesis on the ribosome and can be modulated by the ribosome itself. Such perturbations are generally believed to be mediated through interactions between the nascent chain and the ribosome surface, but despite recent progress in characterising inte...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8513902/ https://www.ncbi.nlm.nih.gov/pubmed/34745542 http://dx.doi.org/10.1039/d1sc04313g |
_version_ | 1784583291920187392 |
---|---|
author | Burridge, Charles Waudby, Christopher A. Włodarski, Tomasz Cassaignau, Anaïs M. E. Cabrita, Lisa D. Christodoulou, John |
author_facet | Burridge, Charles Waudby, Christopher A. Włodarski, Tomasz Cassaignau, Anaïs M. E. Cabrita, Lisa D. Christodoulou, John |
author_sort | Burridge, Charles |
collection | PubMed |
description | The folding of many proteins can begin during biosynthesis on the ribosome and can be modulated by the ribosome itself. Such perturbations are generally believed to be mediated through interactions between the nascent chain and the ribosome surface, but despite recent progress in characterising interactions of unfolded states with the ribosome, and their impact on the initiation of co-translational folding, a complete quantitative analysis of interactions across both folded and unfolded states of a nascent chain has yet to be realised. Here we apply solution-state NMR spectroscopy to measure transverse proton relaxation rates for methyl groups in folded ribosome–nascent chain complexes of the FLN5 filamin domain. We observe substantial increases in relaxation rates for the nascent chain relative to the isolated domain, which can be related to changes in effective rotational correlation times using measurements of relaxation and cross-correlated relaxation in the isolated domain. Using this approach, we can identify interactions between the nascent chain and the ribosome surface, driven predominantly by electrostatics, and by measuring the change in these interactions as the subsequent FLN6 domain emerges, we may deduce their impact on the free energy landscapes associated with the co-translational folding process. |
format | Online Article Text |
id | pubmed-8513902 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-85139022021-11-04 Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy Burridge, Charles Waudby, Christopher A. Włodarski, Tomasz Cassaignau, Anaïs M. E. Cabrita, Lisa D. Christodoulou, John Chem Sci Chemistry The folding of many proteins can begin during biosynthesis on the ribosome and can be modulated by the ribosome itself. Such perturbations are generally believed to be mediated through interactions between the nascent chain and the ribosome surface, but despite recent progress in characterising interactions of unfolded states with the ribosome, and their impact on the initiation of co-translational folding, a complete quantitative analysis of interactions across both folded and unfolded states of a nascent chain has yet to be realised. Here we apply solution-state NMR spectroscopy to measure transverse proton relaxation rates for methyl groups in folded ribosome–nascent chain complexes of the FLN5 filamin domain. We observe substantial increases in relaxation rates for the nascent chain relative to the isolated domain, which can be related to changes in effective rotational correlation times using measurements of relaxation and cross-correlated relaxation in the isolated domain. Using this approach, we can identify interactions between the nascent chain and the ribosome surface, driven predominantly by electrostatics, and by measuring the change in these interactions as the subsequent FLN6 domain emerges, we may deduce their impact on the free energy landscapes associated with the co-translational folding process. The Royal Society of Chemistry 2021-09-09 /pmc/articles/PMC8513902/ /pubmed/34745542 http://dx.doi.org/10.1039/d1sc04313g Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/ |
spellingShingle | Chemistry Burridge, Charles Waudby, Christopher A. Włodarski, Tomasz Cassaignau, Anaïs M. E. Cabrita, Lisa D. Christodoulou, John Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy |
title | Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy |
title_full | Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy |
title_fullStr | Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy |
title_full_unstemmed | Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy |
title_short | Nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by NMR spectroscopy |
title_sort | nascent chain dynamics and ribosome interactions within folded ribosome–nascent chain complexes observed by nmr spectroscopy |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8513902/ https://www.ncbi.nlm.nih.gov/pubmed/34745542 http://dx.doi.org/10.1039/d1sc04313g |
work_keys_str_mv | AT burridgecharles nascentchaindynamicsandribosomeinteractionswithinfoldedribosomenascentchaincomplexesobservedbynmrspectroscopy AT waudbychristophera nascentchaindynamicsandribosomeinteractionswithinfoldedribosomenascentchaincomplexesobservedbynmrspectroscopy AT włodarskitomasz nascentchaindynamicsandribosomeinteractionswithinfoldedribosomenascentchaincomplexesobservedbynmrspectroscopy AT cassaignauanaisme nascentchaindynamicsandribosomeinteractionswithinfoldedribosomenascentchaincomplexesobservedbynmrspectroscopy AT cabritalisad nascentchaindynamicsandribosomeinteractionswithinfoldedribosomenascentchaincomplexesobservedbynmrspectroscopy AT christodouloujohn nascentchaindynamicsandribosomeinteractionswithinfoldedribosomenascentchaincomplexesobservedbynmrspectroscopy |