Action-FRET of a Gaseous Protein

作者：Daly Steven; Knight Geoffrey; Halim Mohamed Abdul; Kulesza Alexander; Choi Chang Min; Chirot Fabien; MacAleese Luke; Antoine Rodolphe; Dugourd Philippe^*

来源：Journal of the American Society for Mass Spectrometry, 2017, 28(1): 38-49.

DOI：10.1007/s13361-016-1449-2

• FRET
• Ubiquitin
• Molecular dynamics
• Action FRET

Mass spectrometry is an extremely powerful technique for analysis of biological molecules, in particular proteins. One aspect that has been contentious is how much native solution-phase structure is preserved upon transposition to the gas phase by soft ionization methods such as electrospray ionization. To address this question-and thus further develop mass spectrometry as a tool for structural biology-structure-sensitive techniques must be developed to probe the gas-phase conformations of proteins. Here, we report Forster resonance energy transfer (FRET) measurements on a ubiquitin mutant using specific photofragmentation as a reporter of the FRET efficiency. The FRET data is interpreted in the context of circular dichroism, molecular dynamics simulation, and ion mobility data. Both the dependence of the FRET efficiency on the charge state-where a systematic decrease is observed-and on methanol concentration are considered. In the latter case, a decrease in FRET efficiency with methanol concentration is taken as evidence that the conformational ensemble of gaseous protein cations retains a memory of the solution phase conformational ensemble upon electrospray ionization.

• 出版日期2017-1