Mapping and identification of Corynebacterium glutamicum proteins by two-dimensional gel electrophoresis and microsequencing

Autor(en): Hermann, T
Wersch, G
Uhlemann, EM
Schmid, R
Burkovski, A
Stichwörter: 2-DIMENSIONAL ELECTROPHORESIS; ACID; BACILLUS-SUBTILIS; Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry; Chemistry, Analytical; Corynebacterium glutamicum; ESCHERICHIA-COLI; IMMOBILIZED PH GRADIENTS; microsequencing; PHYSIOLOGY; protein mapping; proteomics; STATE; two-dimensional polyacrylamide gel electrophoresis
Erscheinungsdatum: 1998
Volumen: 19
Ausgabe: 18
Startseite: 3217
Seitenende: 3221
As a prerequisite for proteome analyses of Corynebacterium glutamicum separation of the cytoplasm and the membrane fraction was optimized and two-dimensional (2-D) gel electrophoresis was established. The resulting 2-D protein maps revealed over 1000 silver-stained protein spots separated by isoelectric point and molecular mass for cytoplasmic proteins and approximately 700 silver-stained spots for proteins of the membrane fraction. Proposing a mean size of 1 kbp per gene the complete C. glutamicum genome of 3 Mbp encodes 3000 different proteins; more than half of these can be located using the maps which are presently available. In this study 10 proteins were identified by N-terminal microsequencing, namely the 35 kDa antigen, antigen 84, ATP synthase subunits alpha, gamma and delta, cysteine synthase, elongation factor G and Ts, enolase, and rotamase. For seven sequences, corresponding proteins could not be identified. Additionally, two proteins were specifically detected by immunoblotting, a corynebacterial porin and the cytoplasmic protein threonine dehydratase. The methods and 2-D maps established in this study will be the basis for comparative studies of protein expression and a detailed proteome analysis of C. glutamicum.
ISSN: 01730835
DOI: 10.1002/elps.1150191827

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