UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

NMR Reveals Functionally Relevant Thermally Induced Structural Changes within the Native Ensemble of G-CSF

Kellerman, Mark-Adam W; Almeida, Teresa; Rudd, Timothy R; Matejtschuk, Paul; Dalby, Paul A; (2022) NMR Reveals Functionally Relevant Thermally Induced Structural Changes within the Native Ensemble of G-CSF. Molecular Pharmaceutics 10.1021/acs.molpharmaceut.2c00398. (In press). Green open access

[thumbnail of acs.molpharmaceut.2c00398.pdf]
Preview
Text
acs.molpharmaceut.2c00398.pdf - Published Version

Download (6MB) | Preview

Abstract

Structure-function relationships in proteins refer to a trade-off between stability and bioactivity, molded by evolution of the molecule. Identifying which protein amino acid residues jeopardize global or local stability for the benefit of bioactivity would reveal residues pivotal to this structure-function trade-off. Here, we use 15N-1H heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) spectroscopy to probe the microenvironment and dynamics of residues in granulocyte colony-stimulating factor (G-CSF) through thermal perturbation. From this analysis, we identified four residues (G4, A6, T133, and Q134) that we classed as significant to global stability, given that they all experienced large environmental and dynamic changes and were closely correlated to each other in their NMR characteristics. Additionally, we observe that roughly four structural clusters are subject to localized conformational changes or partial unfolding prior to global unfolding at higher temperature. Combining NMR observables with structure relaxation methods reveals that these structural clusters concentrate around loop AB (binding site III inclusive). This loop has been previously implicated in conformational changes that result in an aggregation prone state of G-CSF. Residues H43, V48, and S63 appear to be pivotal to an opening motion of loop AB, a change that is possibly also important for function. Hence, we present here an approach to profiling residues in order to highlight their potential roles in the two vital characteristics of proteins: stability and bioactivity.

Type: Article
Title: NMR Reveals Functionally Relevant Thermally Induced Structural Changes within the Native Ensemble of G-CSF
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1021/acs.molpharmaceut.2c00398
Publisher version: https://doi.org/10.1021/acs.molpharmaceut.2c00398
Language: English
Additional information: This is an Open Access article published under a Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/).
Keywords: aggregation, denaturation, remodeling, structure−function, “switch”
UCL classification: UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL
URI: https://discovery.ucl.ac.uk/id/eprint/10154024
Downloads since deposit
40Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item