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Mapping and identification of Corynebacterium glutamicum proteins by two‐dimensional gel electrophoresis and microsequencing
Author(s) -
Hermann Thomas,
Wersch Gregor,
Uhlemann EvaMaria,
Schmid Roland,
Burkovski Andreas
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150191827
Subject(s) - corynebacterium glutamicum , proteome , biology , isoelectric focusing , biochemistry , microbiology and biotechnology , gel electrophoresis , peptide mass fingerprinting , two dimensional gel electrophoresis , proteomics , gene , enzyme
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 α, γ and δ, 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 .