Naslov
Automated Data Reduction for Hydrogen/Deuterium Exchange Experiments

Autori
Kazazić, Saša

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
27th Informal Meeting on Mass Spectrometry 2009 / Allmaier, Guenter ; Marchetti-Deschmann ; Traldi, Pietro ; Vekey, Karoly - Beč : Vienna University of Technology, 2009, 77-77

ISBN
978-3-200-01508

Skup
27th IMMS 2009, Informal Meeting on Mass Spectrometry

Mjesto i datum
Retz, Austrija, 03.05.2009. - 07.05.2009

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
H/D exchange; FT-ICR mass spectrometry; automated data reduction

Sažetak
Mass analysis of proteolytic fragment peptides following hydrogen/deuterium exchange offers a particularly direct and general measure of solvent accessibility (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 interfering species. Applied data reduction procedure could be divided in four major steps. 1) From all raw files centroid m/z value - intensity pairs are extracted applying appropriate threshold cutoff. 2) By analysis of zero exchange time control experiment data, list of all detected peptides with their charge state is generated. 3) Further reduction is made by selection of peptides for which amino acid sequence assignment was possible. 4) For each selected peptide, abundance weighted average m/z value is calculated based on extracted values for isotopic envelope peaks form cumulative data (step 1). The present automated data analysis program interpreted the entire myoglobin HDX data set in ~1.5 h, a vast improvement over manual interpretation requiring weeks of analysis. Errors in data analysis are also reduced. Moreover, the data can be quickly re- analyzed with different input parameters (e.g., different S/N threshold). The algorithm takes full advantage of high mass resolution in analyzing H/D exchange experiments, and uses a novel method to calculate position and width for each nominal m/z of each peptide. In summary, the analysis program permits much greater speed, reliability, and flexibility for analysis of HDX mass spectral data, and thus promises to extend significantly the range of applications of the HDX method.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

POVEZANOST RADA