Enrichment and proteome analysis of a hyperthermostable protein set of archaeon Thermococcus onnurineus NA1

• Published in: Extremophiles : life under extreme conditions Vol. 15; no. 4; pp. 451 - 461
• Main Authors: Yun, Sung-Ho, Choi, Chi-Won, Kwon, Sang Oh, Lee, Yeol Gyun, Chung, Young-Ho, Jung, Hoi Jong, Kim, Yun-Jae, Lee, Jung-Hyun, Choi, Jong-Soon, Kim, Soohyun, Kim, Seung Il
• Format: Journal Article
• Language: English
• Published: Japan Springer Japan 01-07-2011; Springer; Springer Nature B.V
• Subjects: Animal, plant and microbial ecology; Archaeal Proteins - genetics; Archaeal Proteins - metabolism; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biophysics; Biotechnology; E coli; Enzymes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Heat treating; Heat treatment; Hot Temperature; Life Sciences; Mass spectrometry; Metabolism; Microbial Ecology; Microbiology; Original Paper; Protein Stability; Proteins; Proteome - genetics; Proteome - metabolism; Proteomics; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Space life sciences; Thermococcus; Thermococcus - genetics; Thermococcus - metabolism; Thermodynamics; Transition temperatures; Proteome; Archaea; Hyperthermostable; Thermococcales; Archaeobacteria; Bacteria; Thermococcus; Thermococcaceae; Extreme environment; Protein
• ISSN: 1431-0651; 1433-4909
• DOI: 10.1007/s00792-011-0376-1

Thermococcus onnurineus
NA1 is a hyperthermophilic archaeon that can be used for the screening of thermophilic enzymes. Previously, we characterized the metabolic enzymes of the cytosolic proteome by two-dimensional electrophoresis/tandem mass spectrometry (2-DE/MS–MS). In this study, we identified a subset of hyperthermostable proteins in the cytosolic proteome using enrichment by in vitro heat treatment and protein identification. After heat treatment at 100°C for 2 h, 13 and 149 proteins were identified from the soluble proteome subset by 2-DE/MS–MS and 1-DE/MS–MS analysis, respectively. Representative proteins included intracellular protease I, thioredoxin reductase, triosephosphate isomerase, putative hydroperoxide reductase, proteasome, and translation initiation factors. Intracellular protease, deblocking aminopeptidases, and fructose-1,6-bisphosphatase were overexpressed in
Escherichia coli
and biological activity above 85°C was confirmed. The folding transition temperature (Tm) of identified proteins was analyzed using the in silico prediction program TargetStar. The proteins enriched with the heat treatment have higher Tm than the homologous proteins from mesophilic strains. These results suggested that the heat-stable protein set of hyperthermophilic
T. onnurineus
NA1 can be effectively fractionated and enriched by in vitro heat treatment.