Enrichment and proteome analysis of a hyperthermostable protein set of archaeon Thermococcus onnurineus NA1
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Title
- Enrichment and proteome analysis of a hyperthermostable protein set of archaeon Thermococcus onnurineus NA1
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Author(s)
- 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
- KIOST Author(s)
- Kim, Yun Jae(김윤재); Lee, Jung Hyun(이정현)
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Alternative Author(s)
- 정회종; 김윤재; 이정현
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Publication Year
- 2011-07
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Abstract
- 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 100A degrees 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 85A degrees 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.
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ISSN
- 1431-0651
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URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/3848
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DOI
- 10.1007/s00792-011-0376-1
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Bibliographic Citation
- EXTREMOPHILES, v.15, no.4, pp.451 - 461, 2011
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Publisher
- SPRINGER JAPAN KK
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Subject
- COMPLETE GENOME SEQUENCE; HYPERTHERMOPHILIC ARCHAEON; THERMOANAEROBACTER-TENGCONGENSIS; BIOCHEMICAL-CHARACTERIZATION; PYROCOCCUS-FURIOSUS; FRUCTOSE-1,6-BISPHOSPHATASE; THERMOSTABILITY; CRENARCHAEON; PURIFICATION
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Keywords
- Archaea; Thermococcus; Hyperthermostable; Proteome
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Type
- Article
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Language
- English
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Document Type
- Article
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