4.5 Article Proceedings Paper

Comparison of the proteome of Methylobacterium extorquens AM1 grown under methylotrophic and nonmethylotrophic conditions

期刊

PROTEOMICS
卷 4, 期 5, 页码 1247-1264

出版社

WILEY
DOI: 10.1002/pmic.200300713

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C-1 metabolism; matrix-assisted laser desorption/ionization-time of flight mass spectrometry; methylotrophic bacteria; protein identification; two-dimensional gel electrophoresis

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Methylobacterium extorquens AM1 is a facultative methylotrophic bacterium that is capable of growing in the presence of methanol as the sole carbon and energy source, but is also able to grow on a limited number of C-2, C-3, and C-4 compounds, for example succinate. This study provides a proteomic view of the cellular adaptation of M. extorquens AM 1 to growth on methanol and succinate, respectively. Cytosolic proteins were separated by two-dimensional gel electrophoresis employing overlapping pH ranges and visualized by silver nitrate or fluorescence staining. A proteomic reference map containing 229 different proteins identified by peptide mass fingerprinting of tryptic fragments was established. Comparative proteome profiling of methanol- and succinate-grown cells led to the identification of 68 proteins that are induced under methylotrophic growth conditions in comparison to growth on succinate. This group includes most proteins known to be directly involved in methanol oxidation to CO2 and in assimilation of one carbon units by the serine cycle as well as 18 proteins without any assigned function and two proteins with a predicted regulatory function. Furthermore, the proteome analysis revealed putative isoenzymes for formaldehyde-activating enzyme Fae, malyl-CoA lyase, malate-dehydrogenase, and fumarase, that need to be characterized functionally in future studies.

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