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. 2007 Oct;389(4):1055-64. doi: 10.1007/s00216-006-0997-x. Epub 2007 Jan 13.

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/Fe(3+) siderophore transport system, the maltose binding protein, and a subunit of the cytochrome bc-aa(3) 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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / metabolism
  • Bacterial Proteins / metabolism
  • Betaine / analogs & derivatives
  • Betaine / chemistry
  • Chromatography, Ion Exchange / methods
  • Corynebacterium glutamicum / chemistry
  • Corynebacterium glutamicum / genetics
  • Corynebacterium glutamicum / metabolism*
  • Electron Transport Chain Complex Proteins / metabolism
  • Electrophoresis, Polyacrylamide Gel / methods
  • Fatty Alcohols
  • Genotype
  • Lysine / biosynthesis*
  • Membrane Proteins / analysis
  • Membrane Proteins / metabolism*
  • Models, Biological
  • Proteome / analysis
  • Proteome / metabolism*
  • Proteomics / methods*
  • Quaternary Ammonium Compounds / chemistry
  • Reproducibility of Results
  • Succinate Dehydrogenase / metabolism
  • Up-Regulation

Substances

  • ATP-Binding Cassette Transporters
  • Bacterial Proteins
  • Electron Transport Chain Complex Proteins
  • Fatty Alcohols
  • Membrane Proteins
  • Proteome
  • Quaternary Ammonium Compounds
  • amidosulfobetaine-14
  • Betaine
  • cetalkonium chloride
  • Succinate Dehydrogenase
  • Lysine