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The use of recently developed mass spectrometry-based proteomic approaches for the study of methylocella silvestris BL2
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Patel, Nisha A. (2012) The use of recently developed mass spectrometry-based proteomic approaches for the study of methylocella silvestris BL2. PhD thesis, University of Warwick.
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WRAP_THESIS_Patel_2012.pdf - Submitted Version Download (16Mb) | Preview |
Official URL: http://webcat.warwick.ac.uk/record=b2680503~S1
Abstract
The study of the protein complement, termed proteomics, has advanced over the last
twenty years as a consequence of developments in mass spectrometry. Currently,
improvements in mass spectrometry-based approaches are targeted towards achieving
information on both the identity and abundance of proteins.
Increased numbers of protein identifications are obtained by simplifying the
analyte of interest. This can be achieved with the use of separation techniques,
including two-dimensional liquid chromatography (2D-LC). Ion mobility coupled to
mass spectrometry has recently been shown to be a useful post-ionisation separation
tool for proteomic studies. The utility of these technologies for obtaining both
qualitative and quantitative information is not extensively addressed in the current
literature.
The use of a recently developed 2D-LC system, together with a method of ion
mobility separation and a label-free quantitative approach for proteomic studies has
been evaluated here for characterising the proteome of the bacterium Methylocella
silvestris. This bacterium is the first methane-utilising bacteria also discovered to
grow on substrates containing carbon-carbon bonds, and has great biotechnological
potential. The metabolism of this bacterium was studied by obtaining information on
its soluble proteome when grown with methane, propane, succinate, acetate,
methanol, methylamine or trimethylamine.
The benefits and limitations of 2D-LC and ion mobility for profiling and labelfree
quantitative studies were demonstrated for simple mixtures and complex bacterial
extracts. The combination of both 2D-LC and ion mobility was also achieved,
resulting in wider proteome coverage when compared to the respective stand-alone
approaches.
A cluster of expressed genes that were greatly up-regulated under
trimethylamine growth and monomethylamine growth were proposed to be involved
in the indirect pathway for trimethylamine metabolism. It was further verified that
one of these genes expresses the previously unidentified trimethylamine
monooxygenase. A propane assimilation route was proposed, based on information
obtained on the levels of primary oxidation enzymes and downstream central
metabolic pathways.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QD Chemistry Q Science > QP Physiology Q Science > QR Microbiology |
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Library of Congress Subject Headings (LCSH): | Mass spectrometry, Gram-negative bacteria, Liquid chromatography, Proteomics | ||||
Official Date: | September 2012 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Scrivens, James H.; Slade, Susan E. | ||||
Sponsors: | Engineering and Physical Sciences Research Council (EPSRC); Royal Society of Chemistry (Great Britain) (RSC) | ||||
Extent: | xviii, 212 leaves : illustrations, charts. | ||||
Language: | eng |
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