Automated data reduction for hydrogen/deuterium exchange experiments, enabled by high-resolution Fourier transform ion cyclotron resonance mass spectrometry (CROSBI ID 157704)
Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija
Podaci o odgovornosti
Kazazić, Saša ; Zhang, Hui-Min ; Schaub, Tanner M. ; Emmett, Mark R. ; Hendrickson, Christopher L. ; Blakney, Gregory T. ; Marshall, Alan G.
engleski
Automated data reduction for hydrogen/deuterium exchange experiments, enabled by high-resolution Fourier transform ion cyclotron resonance mass spectrometry
Mass analysis of proteolytic fragment peptides following hydrogen/deuterium exchange offers a general measure of solvent accessibility/hydrogen bonding (and thus conformation) of solution-phase proteins and their complexes. The primary problem in such mass analyses is reliable and rapid assignment of mass spectral peaks to the correct charge state and degree of deuteration of each fragment peptide, in the presence of substantial overlap between isotopic distributions of target peptides, autolysis products, and other interferant species. Here, we show that at sufficiently high mass resolving power (m/delta m50% > 100, 000), it becomes possible to resolve enough of those overlaps that automated data reduction becomes possible, based on the actual elemental composition of each peptide without the need to deconvolve isotopic distributions. We demonstrate automated, rapid, reliable assignment of peptide masses from H/D exchange experiments, based on electrospray ionization FT-ICR mass spectra from H/D exchange of solution-phase myoglobin. Combined with previously demonstrated automated data acquisition for such experiments, the present data reduction algorithm enhances automation (and thus expanded generality and applicability) for high-resolution mass spectrometry-based analysis of H/D exchange of solution-phase proteins.
peptide ; protein ; H/D exchange ; autosampler ; mass spectrometry ; ICR ; ESI ; FT-ICR ; FTMS ; LTQFT
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Podaci o izdanju
21 (4)
2010.
550-558
objavljeno
1044-0305
1879-1123
10.1016/j.jasms.2009.12.016