Patel, Nisha A., Crombie, Andrew, Slade, Susan E., Thalassinos, Konstantinos, Hughes, Chris, Connolly, Joanne B., Langridge, James, Murrell, J. Colin and Scrivens, James H.. (2012) Comparison of one- and two-dimensional liquid chromatography approaches in the label-free quantitative analysis of methylocella silvestris. Journal of Proteome Research, Vol.11 (No.9). pp. 4755-4763. ISSN 1535-3893

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Abstract

The proteome of the bacterium Methylocella silvestris has been characterized using reversed phase ultra high pressure liquid chromatography (UPLC) and two-dimensional reversed phase (high pH)–reversed phase (low pH) UPLC prior to mass spectrometric analysis. Variations in protein expression levels were identified with the aid of label-free quantification in a study of soluble protein extracts from the organism grown using methane, succinate, or propane as a substrate. The number of first dimensional fractionation steps has been varied for 2D analyses, and the impact on data throughput and quality has been demonstrated. Comparisons have been made regarding required experimental considerations including total loading of biological samples required, instrument time, and resulting data file sizes. The data obtained have been evaluated with respect to number of protein identifications, confidence of assignments, sequence coverage, relative levels of proteins, and dynamic range. Good qualitative and quantitativ...

Item Type:	Journal Article
Divisions:	Faculty of Science > Life Sciences (2010- )
Journal or Publication Title:	Journal of Proteome Research
Publisher:	American Chemical Society
ISSN:	1535-3893
Date:	2012
Volume:	Vol.11
Number:	No.9
Page Range:	pp. 4755-4763
Identification Number:	10.1021/pr300253s
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
URI:	http://wrap.warwick.ac.uk/id/eprint/50476

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