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Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis

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....

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Autores principales: Zhang, Huoming, Bensaddek, Dalila
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8469718/
https://www.ncbi.nlm.nih.gov/pubmed/34575131
http://dx.doi.org/10.3390/life11090982
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author Zhang, Huoming
Bensaddek, Dalila
author_facet Zhang, Huoming
Bensaddek, Dalila
author_sort Zhang, Huoming
collection PubMed
description 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 an Arabidopsis sample with spike-in 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 54.7% increase in the number of proteins quantified than with cDIA analysis. This is particularly important for low abundance proteins, as exemplified by the six-protein mix spike-in. In cDIA only five out of the six-protein mix were quantified while our approach allowed accurate quantitation of all six proteins.
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spelling pubmed-84697182021-09-27 Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis Zhang, Huoming Bensaddek, Dalila Life (Basel) Article 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 an Arabidopsis sample with spike-in 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 54.7% increase in the number of proteins quantified than with cDIA analysis. This is particularly important for low abundance proteins, as exemplified by the six-protein mix spike-in. In cDIA only five out of the six-protein mix were quantified while our approach allowed accurate quantitation of all six proteins. MDPI 2021-09-18 /pmc/articles/PMC8469718/ /pubmed/34575131 http://dx.doi.org/10.3390/life11090982 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Huoming
Bensaddek, Dalila
Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis
title Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis
title_full Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis
title_fullStr Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis
title_full_unstemmed Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis
title_short Narrow Precursor Mass Range for DIA–MS Enhances Protein Identification and Quantification in Arabidopsis
title_sort narrow precursor mass range for dia–ms enhances protein identification and quantification in arabidopsis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8469718/
https://www.ncbi.nlm.nih.gov/pubmed/34575131
http://dx.doi.org/10.3390/life11090982
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