Life-style changes of a halophilic archaeon analyzed by quantitative proteomics.
Quantitative proteomics based on isotopic labeling has become the method of choice to accurately determine changes in protein abundance in highly complex mixtures. Isotope-coded protein labeling (ICPL), which is based on the nicotinoylation of proteins at lysine residues and free N-termini was used as a simple, reliable and fast method for the comparative analysis of three different cellular states of the halophilic archaeon Halobacterium salinarum through pairwise comparison. The labeled proteins were subjected to SDS-PAGE, in-gel digested and the proteolytic peptides were separated by LC and analyzed by MALDI-TOF/TOF MS. Automated quantitation was performed by comparing the MS peptide signals of (12)C and (13)C nicotinoylated isotopic peptide pairs. The transitions between (i) aerobic growth in complex versus synthetic medium and (ii) aerobic versus anaerobic/phototrophic growth, both in complex medium, provide a wide span in nutrient and energy supply for the cell and thus allowed optimal studies of proteome changes. In these two studies, 559 and 643 proteins, respectively, could be quantified allowing a detailed analysis of the adaptation of H. salinarum to changes of its living conditions. The subtle cellular response to a wide variation of nutrient and energy supply demonstrates a fine tuning of the cellular protein inventory.
Tebbe
Andreas
A
Department of Membrane Biochemistry, Max Planck Institute of Biochemistry, Martinsried, Germany.
Schmidt
Alexander
A
Konstantinidis
Kosta
K
Falb
Michaela
M
Bisle
Birgit
B
Klein
Christian
C
Aivaliotis
Michalis
M
Kellermann
Josef
J
Siedler
Frank
F
Pfeiffer
Friedhelm
F
Lottspeich
Friedrich
F
Oesterhelt
Dieter
D
D019843:Archaeal Proteins / Q000032:analysis* / Q000378:metabolism; D019617:Halobacterium salinarum / Q000254:growth & development / Q000378:metabolism*; D007553:Isotope Labeling; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D019843:Archaeal Proteins; D020543:Proteome