caa a22 4500 445818638 CHVBK 20180317145236.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s00792-011-0376-1 doi (NATIONALLICENCE)springer-10.1007/s00792-011-0376-1 Enrichment and proteome analysis of a hyperthermostable protein set of archaeon Thermococcus onnurineus NA1 [Elektronische Daten] [Sung-Ho Yun, Chi-Won Choi, Sang Kwon, Yeol Lee, Young-Ho Chung, Hoi Jung, Yun-Jae Kim, Jung-Hyun Lee, Jong-Soon Choi, Soohyun Kim, Seung Kim] 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 2h, 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. Springer, 2011 Archaea nationallicence Thermococcus nationallicence Hyperthermostable nationallicence Proteome nationallicence Yun Sung-Ho Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Choi Chi-Won Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Kwon Sang Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Lee Yeol Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Chung Young-Ho Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Jung Hoi Korea Ocean Research & Development Institute, Ansan, P.O. Box 29, 425-600, Seoul, Korea aut Kim Yun-Jae Korea Ocean Research & Development Institute, Ansan, P.O. Box 29, 425-600, Seoul, Korea aut Lee Jung-Hyun Korea Ocean Research & Development Institute, Ansan, P.O. Box 29, 425-600, Seoul, Korea aut Choi Jong-Soon Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Kim Soohyun Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Kim Seung Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut Extremophiles Springer Japan 15/4(2011-07-01), 451-461 1431-0651 15:4<451 2011 15 792 https://doi.org/10.1007/s00792-011-0376-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-011-0376-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yun Sung-Ho Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Choi Chi-Won Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Kwon Sang Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Lee Yeol Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Chung Young-Ho Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Jung Hoi Korea Ocean Research & Development Institute, Ansan, P.O. Box 29, 425-600, Seoul, Korea aut NATIONALLICENCE 700 1- Kim Yun-Jae Korea Ocean Research & Development Institute, Ansan, P.O. Box 29, 425-600, Seoul, Korea aut NATIONALLICENCE 700 1- Lee Jung-Hyun Korea Ocean Research & Development Institute, Ansan, P.O. Box 29, 425-600, Seoul, Korea aut NATIONALLICENCE 700 1- Choi Jong-Soon Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Kim Soohyun Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 700 1- Kim Seung Division of Life Science, Korea Basic Science Institute, 305-333, Daejeon, Korea aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/4(2011-07-01), 451-461 1431-0651 15:4<451 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer