title: Action-FRET of a Gaseous Protein.
abstract: 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 Förster 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. Graphical Abstract ᅟ.
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Steven
authors: S
authors: Daly
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Geoffrey
authors: G
authors: Knight
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Mohamed Abdul
authors: MA
authors: Halim
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Alexander
authors: A
authors: Kulesza
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Chang Min
authors: CM
authors: Choi
authors: Institut des Sciences Analytiques, Université Lyon 1 - CNRS, Université de Lyon, 69622, Villeurbanne Cedex, France.
authors: Fabien
authors: F
authors: Chirot
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Luke
authors: L
authors: MacAleese
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France.
authors: Rodolphe
authors: R
authors: Antoine
authors: Institut Lumière Matière, Université Lyon 1-CNRS, Université de Lyon, 69622, Villeurbanne cedex, France. philippe.dugourd@univ-lyon1.fr.
authors: Philippe
authors: P
authors: Dugourd
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002412:Cations / Q000737:chemistry; D002417:Cattle; D002942:Circular Dichroism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005740:Gases / Q000737:chemistry; D056004:Molecular Dynamics Simulation; D009154:Mutation; D011487:Protein Conformation; D021241:Spectrometry, Mass, Electrospray Ionization; D055672:Static Electricity; D025801:Ubiquitin / Q000737:chemistry* / Q000235:genetics
chemical_list: D002412:Cations; D005740:Gases; D025801:Ubiquitin
keywords: Action FRET; FRET; Molecular dynamics; Ubiquitin