Fluorescence resonance energy transfer in gaseous, mass-selected polyproline peptides.
Despite the many successes of mass spectrometry in the analysis of biological samples, the need to better understand the correlation between condensed-phase properties and those of electrospray species remains. In particular, the link between structures in the condensed phase and in the gaseous environment of the mass spectrometer is still elusive. Here, we show that fluorescence resonance energy transfer (FRET) can be used to probe the conformations of gaseous biopolymers which are formed by electrospray ionization (ESI) and manipulated in a quadrupole ion trap mass spectrometer. A rhodamine dye pair suitable for gas-phase FRET is characterized. Both steady state spectra and lifetime measurements are used to monitor energy transfer in a series of dye-labeled polyproline-based peptides. FRET efficiency is explored as a function of peptide chain length and charge state. For the peptide with eight proline repeats, virtually complete energy transfer is observed. For the peptide with 14 proline repeats, energy transfer decreases as the charge state increases, consistent with Coulomb repulsion induced elongation of the peptide backbone. FRET measurements of the longest peptide examined, which has 20 proline repeats, indicates that the peptide adopts a bent configuration. Evidence for multiple conformations present within the ensemble of trapped ions is provided by fluorescence lifetime measurements. Gas-phase FRET measurements promise to be a new route to probe the conformations of large gaseous ions.
Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada.
Francis O
FO
Talbot
Anthony
A
Rullo
Huihui
H
Yao
Rebecca A
RA
Jockusch
D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005740:Gases / Q000737:chemistry; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation
D005740:Gases; D010455:Peptides; C011083:polyproline