Evaluation of two proteomics technologies used to screen the membrane proteomes of wild-type Corynebacterium glutamicum and an L-lysine-producing strain.
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.
Schluesener
Daniela
D
Lehrstuhl für Biochemie der Pflanzen, Ruhr Universität Bochum, 44780, Bochum, Germany.
Rögner
Matthias
M
Poetsch
Ansgar
A
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001622:Betaine / Q000031:analogs & derivatives / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000379:methods; D048230:Corynebacterium glutamicum / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045222:Electron Transport Chain Complex Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005233:Fatty Alcohols; D005838:Genotype; D008239:Lysine / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D008954:Models, Biological; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D015203:Reproducibility of Results; D013385:Succinate Dehydrogenase / Q000378:metabolism; D015854:Up-Regulation
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D045222:Electron Transport Chain Complex Proteins; D005233:Fatty Alcohols; D008565:Membrane Proteins; D020543:Proteome; D000644:Quaternary Ammonium Compounds; C506587:amidosulfobetaine-14; D001622:Betaine; C012817:cetalkonium chloride; D013385:Succinate Dehydrogenase; D008239:Lysine