Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry
Abstract Understanding the effect of temperature to the structural integrity of proteins is relevant to diverse areas such as biotechnology and climate change. Variable temperature ion mobility-mass spectrometry (VT-IM-MS) can measure the effect of temperature on conformational landscapes. To deline...
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| Format: | Article |
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Nature Portfolio
2025-05-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-59065-x |
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| author | Xudong Wang Emma Norgate Junxiao Dai Florian Benoit Tony Bristow Richard M. England Jason M. D. Kalapothakis Perdita E. Barran |
| author_facet | Xudong Wang Emma Norgate Junxiao Dai Florian Benoit Tony Bristow Richard M. England Jason M. D. Kalapothakis Perdita E. Barran |
| author_sort | Xudong Wang |
| collection | DOAJ |
| description | Abstract Understanding the effect of temperature to the structural integrity of proteins is relevant to diverse areas such as biotechnology and climate change. Variable temperature ion mobility-mass spectrometry (VT-IM-MS) can measure the effect of temperature on conformational landscapes. To delineate collision effects from structural change we report measurements using molecules with different degrees of rigidity namely: poly (L-lysine) (PLL) dendrimer, ubiquitin, β-casein and α-synuclein from 190-350 K. The CCS of PLL dendrimer varies with temperature consistent with collision theory, by contrast, the structure of each protein alters with notable restructuring at 350 K and 250 K, following predicted in vitro stability curves. At 210 K and 190 K we kinetically trap unfolding intermediates. For alpha-synuclein, the 13+ ions present two distinct conformers and VT-IM-MS measurements allow us to calculate the transition rate and activation energies of their conversion. These data exemplify the capacity of VT-IM-MS to provide insights on the thermodynamics of conformational restructuring. |
| format | Article |
| id | doaj-art-e00e03939ac24fb4aed2812116dd0032 |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-e00e03939ac24fb4aed2812116dd00322025-08-20T03:09:20ZengNature PortfolioNature Communications2041-17232025-05-0116111310.1038/s41467-025-59065-xConformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometryXudong Wang0Emma Norgate1Junxiao Dai2Florian Benoit3Tony Bristow4Richard M. England5Jason M. D. Kalapothakis6Perdita E. Barran7Michael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of ManchesterMichael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of ManchesterMichael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of ManchesterMichael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of ManchesterChemical Development, Pharmaceutical Technology and Development, Operations, AstraZenecaAdvanced Drug Delivery, Pharmaceutical Sciences, R&D, AstraZenecaMichael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of ManchesterMichael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of ManchesterAbstract Understanding the effect of temperature to the structural integrity of proteins is relevant to diverse areas such as biotechnology and climate change. Variable temperature ion mobility-mass spectrometry (VT-IM-MS) can measure the effect of temperature on conformational landscapes. To delineate collision effects from structural change we report measurements using molecules with different degrees of rigidity namely: poly (L-lysine) (PLL) dendrimer, ubiquitin, β-casein and α-synuclein from 190-350 K. The CCS of PLL dendrimer varies with temperature consistent with collision theory, by contrast, the structure of each protein alters with notable restructuring at 350 K and 250 K, following predicted in vitro stability curves. At 210 K and 190 K we kinetically trap unfolding intermediates. For alpha-synuclein, the 13+ ions present two distinct conformers and VT-IM-MS measurements allow us to calculate the transition rate and activation energies of their conversion. These data exemplify the capacity of VT-IM-MS to provide insights on the thermodynamics of conformational restructuring.https://doi.org/10.1038/s41467-025-59065-x |
| spellingShingle | Xudong Wang Emma Norgate Junxiao Dai Florian Benoit Tony Bristow Richard M. England Jason M. D. Kalapothakis Perdita E. Barran Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry Nature Communications |
| title | Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry |
| title_full | Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry |
| title_fullStr | Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry |
| title_full_unstemmed | Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry |
| title_short | Conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility-mass spectrometry |
| title_sort | conformational landscapes of rigid and flexible molecules explored with variable temperature ion mobility mass spectrometry |
| url | https://doi.org/10.1038/s41467-025-59065-x |
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