Proteomic characterization of the sulfur-reducing hyperthermophilic archaeon Thermococcus onnurineus NA1 by 2-DE/MS-MS.
Thermococcus onnurineus NA1, a sulfur-reducing hyperthermophilic archaeon, was isolated from a deep-sea hydrothermal vent area in Papua New Guinea. The strain requires elemental sulfur as a terminal electron acceptor for heterotrophic growth on peptides, amino acids and sugars. Recently, genome sequencing of Thermococcus onnurineus NA1 was completed. In this study, 2-DE/MS-MS analysis of the cytosolic proteome was performed to elucidate the metabolic characterization of Thermococcus onnurineus NA1 at the protein level. Among the 1,136 visualized protein spots, 110 proteins were identified. Enzymes related to metabolic pathways of amino acids utilization, glycolysis, pyruvate conversion, ATP synthesis, and protein synthesis were identified as abundant proteins, highlighting the fact that these are major metabolic pathways in Thermococcus onnurineus NA1. Interestingly, multiple spots of phosphoenolpyruvate synthetase and elongation factor Tu were found on 2D gels generated by truncation at the N-terminus, implicating the cellular regulatory mechanism of this key enzyme by protease degradation. In addition to the proteins involved in metabolic systems, we also identified various proteases and stress-related proteins. The proteomic characterization of abundantly induced proteins using 2-DE/MS-MS enables a better understanding of Thermococcus onnurineus NA1 metabolism.
Kwon
Sang Oh
SO
Korea Basic Science Institute, Daejeon 305-333, South Korea.
Kang
Sung Gyun
SG
Park
Soon-Ho
SH
Kim
Young Hwan
YH
Choi
Jong-Soon
JS
Lee
Jung-Hyun
JH
Kim
Seung Il
SI
D001105:Archaea / Q000235:genetics / Q000378:metabolism; D019843:Archaeal Proteins / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006019:Glycolysis; D013058:Mass Spectrometry / Q000379:methods*; D008954:Models, Biological; D010219:Papua New Guinea; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D013455:Sulfur / Q000737:chemistry*; D019712:Thermococcus / Q000378:metabolism*; D014871:Water Microbiology
D019843:Archaeal Proteins; D020543:Proteome; D013455:Sulfur