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Evaluation of two proteomics technologies used to screen the membrane proteomes of wild-type Corynebacterium glutamicum and an L-lysine-producing strain

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Abstract

The membrane proteomes of a wild-type Corynebacterium glutamicum and an L-lysine-producing strain were quantitatively analyzed by two complementary proteomics techniques—anion exchange chromatography AIEC/SDS-PAGE and 16BAC-PAGE/SDS-PAGE—and the results were compared. Although both techniques allow for the fast screening of differences in protein abundance, AIEC/SDS-PAGE was superior to 16BAC-PAGE/SDS-PAGE with respect to protein separation, it was more suitable for relative protein quantification, and allowed more differentially regulated proteins to be detected (the succinate dehydrogenase complex, an ABC-type cobalamin/Fe3+ siderophore transport system, the maltose binding protein, and a subunit of the cytochrome bc-aa3 supercomplex were upregulated, while a periplasmic component of an ABC-type transporter and an iron-regulated ABC-type transporter were downregulated in the producer). The results indicate the important role of tricarboxylic acid cycle enzymes as well as the adaptation of transport processes in L-lysine-producing cells. Since the only genetic differences between the wild type and the L-lysine producer occur between four central metabolic enzymes in the cytoplasm, our study illustrates the complex effects of metabolic engineering on cell physiology and the power of the new AIEC/SDS-PAGE proteomics approach to detect these effects.

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Abbreviations

AIEC:

anion exchange chromatography

ASB-14:

amidosulfobetaine-14

16BAC:

benzyldimethyl-n-hexadecylammoniumchloride

TM:

transmembrane helix

MALDI-TOF PMF:

matrix-assisted laser desorption/ionization time-of-flight peptide mass fingerprinting

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Acknowledgements

The authors would like to thank Ursula Hilp for excellent technical assistance. This work was supported by grants from the Bundesministerium für Bildung und Forschung (BMBF 0312843E), the Deutsche Forschungsgemeinschaft (European Graduate College 795) and by the Fonds der Chemischen Industrie.

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Authors and Affiliations

  1. Lehrstuhl für Biochemie der Pflanzen, Ruhr Universität Bochum, 44780, Bochum, Germany

    Daniela Schluesener, Matthias Rögner & Ansgar Poetsch

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  1. Daniela Schluesener
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  2. Matthias Rögner
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  3. Ansgar Poetsch
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Correspondence to Ansgar Poetsch.

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Schluesener, D., Rögner, M. & Poetsch, A. Evaluation of two proteomics technologies used to screen the membrane proteomes of wild-type Corynebacterium glutamicum and an L-lysine-producing strain. Anal Bioanal Chem 389, 1055–1064 (2007). https://doi.org/10.1007/s00216-006-0997-x

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  • DOI: https://doi.org/10.1007/s00216-006-0997-x

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