Zhang, H.; Bensaddek, D. Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis. Life2021, 11, 982.
Zhang, H.; Bensaddek, D. Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis. Life 2021, 11, 982.
Zhang, H.; Bensaddek, D. Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis. Life2021, 11, 982.
Zhang, H.; Bensaddek, D. Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis. Life 2021, 11, 982.
Abstract
Data independent acquisition - mass spectrometry (DIA-MS) is becoming widely utilised for robust and accurate quantification of samples in quantitative proteomics. Here, we describe the systematic evaluation of the effects of DIA precursor mass range on total protein identification and quantification. We show that a narrow mass range of precursors (~250 m/z) for DIA-MS enables a higher number of protein identifications. Subsequent application of DIA with narrow precursor range (from 400 to 650 m/z) on Arabidopsis sample with spike-in of known proteins identified 34.7% more proteins than in conventional DIA (cDIA) with a wide precursor range of 400-1200 m/z. When combining several DIA-MS analyses with narrow precursor ranges (i.e., 400-650, 650-900 and 900-1200 m/z), we were able to quantify 10,099 protein groups with a median coefficient of variation of <6%. These findings represent a 59.4% increase in the number of proteins quantified than with cDIA analysis. This is particularly important for low abundance proteins, as exemplified by the 6-protein mix spike-in. In cDIA only 5 out of the 6-protein mix were quantified while our approach allowed accurate quantitation of all six proteins.
Keywords
data-independent acquisition (DIA); mass spectrometry, precursor mass range selection, Arabidopsis; quantitative proteomics.
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
