Evaluation of sample preparation methods for mass spectrometry-based proteomic analysis of barley leaves

Plant Methods
Wei-Qing WangAdelina Rogowska-Wrzesinska

Abstract

Sample preparation is a critical process for proteomic studies. Many efficient and reproducible sample preparation methods have been developed for mass spectrometry-based proteomic analysis of human and animal tissues or cells, but no attempt has been made to evaluate these protocols for plants. We here present an LC-MS/MS-based proteomics study of barley leaf aimed at optimization of methods to achieve efficient and unbiased trypsin digestion of proteins prior to LC-MS/MS based sequencing and quantification of peptides. We evaluated two spin filter-aided sample preparation protocols using either sodium dodecyl-sulphate or sodium deoxycholate (SDC), and three in-solution digestion (ISD) protocols using SDC or trichloroacetic acid/acetone precipitation. The proteomics workflow identified and quantified up to 1800 barley proteins based on sequencing of up to 6900 peptides per sample. The two spin filter-based protocols provided a 12-38% higher efficiency than the ISD protocols, including more proteins of low abundance. Among the ISD protocols, a simple one-step reduction and S-alkylation method (OP-ISD) was the most efficient for barley leaf sample preparation; it identified and quantified 1500 proteins and displayed higher pepti...Continue Reading

References

Jul 27, 2002·Analytical Chemistry·Fanyu MengNeil L Kelleher
Mar 12, 2005·Proteomics·Linda L ManzaDaniel C Liebler
Apr 5, 2007·Nature Protocols·Tal IsaacsonJocelyn K C Rose
Dec 12, 2007·Journal of Separation Science·Constantine D Stalikas
Jan 11, 2008·Journal of Proteome Research·Takeshi MasudaYasushi Ishihama
Jul 11, 2008·Journal of Separation Science·Wei WangShaoning Chen
Apr 30, 2009·Annual Review of Biomedical Engineering·John R YatesAleksey Nakorchevsky
Mar 24, 2012·Annual Review of Biochemistry·Ariel BensimonRuedi Aebersold
Jun 25, 2013·Molecular & Cellular Proteomics : MCP·Ileana R LeónRichard R Sprenger
Apr 2, 2015·Journal of Proteomics·Xiao-lin ZhangTian-lai Li
Apr 11, 2015·Nature Reviews. Molecular Cell Biology·Mark Larance, Angus I Lamond
Oct 2, 2015·Methods in Molecular Biology·Peter Højrup
Nov 4, 2015·Nucleic Acids Research·Juan Antonio VizcaínoHenning Hermjakob
Dec 16, 2016·Advances in Experimental Medicine and Biology·Sophie Alvarez, Michael J Naldrett
Jan 24, 2017·The Plant Journal : for Cell and Molecular Biology·Jedrzej SzymanskiAsaph Aharoni

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Citations

Feb 28, 2020·International Journal of Molecular Sciences·Olga A AndrzejczakKim H Hebelstrup
Dec 14, 2019·Plant Methods·Yahui LiuLanping Guo
Dec 7, 2020·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Liting DengPaul A Haynes
Dec 5, 2020·International Journal of Molecular Sciences·Galina SmolikovaAndrej Frolov
Mar 30, 2021·Frontiers in Plant Science·Kamil MikulášekZbyněk Zdráhal
Jul 29, 2021·Journal of Agricultural and Food Chemistry·Mahya BahmaniMichelle L Colgrave
Sep 17, 2021·The Plant Journal : for Cell and Molecular Biology·Ian Max MøllerOlivier Van Aken

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Methods Mentioned

BETA
electrophoresis
acetylation
PCA
laser

Software Mentioned

TMHMM Server
ExPASy
Excel
Proteome Discoverer
RapiGest
Ensemblplant
Qubit

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