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Bottom-up proteomics of envelope proteins extracted from spinach chloroplast via high organic content CE-MS.

Authors :
Cheng J
Morin GB
Chen DDY
Source :
Electrophoresis [Electrophoresis] 2020 Mar; Vol. 41 (5-6), pp. 370-378. Date of Electronic Publication: 2020 Feb 19.
Publication Year :
2020

Abstract

A high organic content CE-MS/MS (HOCE-MS/MS) method was developed for the proteomic analysis of envelope proteins extracted from spinach leaves. Separation was performed in a 1-m long hydroxypropyl cellulose coated capillary, using 8% (v/v) formic acid in 70% (v/v) methanol and 22% water as the BGE. A flow-through microvial interface was used to couple the CE system with an Orbitrap Fusion Lumos mass spectrometer, and field-amplified sample stacking was used to improve the concentration sensitivity. Using this optimized method, 3579 peptides and 1141 proteins were identified using the Proteome Discoverer software with a 1% false discovery rate at the protein level. Relative to conventional aqueous CE, HOCE-MS did a better job of discovering hydrophobic peptides and provided more peptide and protein identifications. Relative to nano-LC-MS, it achieved comparable peptide and protein identification performance and detected peptides not identified by LC-MS: of the full set of peptides identified using the two techniques, 19% were identified only using HOCE-MS. It also outperformed nano-LC-MS with respect to the detection of low molecular weight peptides.<br /> (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Details

Language :
English
ISSN :
1522-2683
Volume :
41
Issue :
5-6
Database :
MEDLINE
Journal :
Electrophoresis
Publication Type :
Academic Journal
Accession number :
31994203
Full Text :
https://doi.org/10.1002/elps.201900452