Wang Xudong, Norgate Emma, Dai Junxiao, Benoit Florian, Bristow Tony, England Richard M, Kalapothakis Jason M D, Barran Perdita E
Michael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
Chemical Development, Pharmaceutical Technology and Development, Operations, AstraZeneca, Charter Way, Macclesfield, SK102NA, UK.
Nat Commun. 2025 May 6;16(1):4183. doi: 10.1038/s41467-025-59065-x.
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.
了解温度对蛋白质结构完整性的影响与生物技术和气候变化等多个领域相关。变温离子淌度-质谱（VT-IM-MS）可以测量温度对构象态势的影响。为了区分碰撞效应和结构变化，我们报告了使用不同刚性程度的分子进行的测量，即：聚（L-赖氨酸）（PLL）树枝状大分子、泛素、β-酪蛋白和α-突触核蛋白，温度范围为190 - 350K。PLL树枝状大分子的截面碰撞面积（CCS）随温度变化，符合碰撞理论，相比之下，每种蛋白质的结构在350K和250K时发生显著重组，符合预测的体外稳定性曲线。在210K和190K时，我们动力学捕获了展开中间体。对于α-突触核蛋白，13 +离子呈现两种不同的构象，VT-IM-MS测量使我们能够计算它们转化的过渡速率和活化能。这些数据例证了VT-IM-MS在提供构象重组热力学见解方面的能力。
