Comparative proteomic analysis of extracellular vesicles (EVs) from the hyperthermophilic archaeon, Thermococcusonnurineus NA1

Title
Comparative proteomic analysis of extracellular vesicles (EVs) from the hyperthermophilic archaeon, Thermococcusonnurineus NA1
Author(s)
최동희; 권용민; 윤성호; 김승일; 배승섭; 강성균; 이정현; 김상진
KIOST Author(s)
Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Alternative Author(s)
최동희; 권용민; 배승섭; 강성균; 이정현; 김상진
Publication Year
2014-05-05
Abstract
Thermococcus onnurineus NA1, a sulfur-reducing hyperthermophilic archaeon, is able to produce extracellular vesicles (ToEVs) under anaerobic cultivation at 80 oC. EVs were purified through ultra membrane filtration and CsCl-density gradient ultracentrifugation. Two major bands (B & C) were obtained and theirs proteomes were further analyzed. ToEVs were spherical in shape and the size varied from 150 nm to 250 nm in diameter, and the vesicles of both band showed identical shape and size. The proteins of these two kinds of ToEVs were separated and identified by one-dimensional SDS-PAGE and the quantitative proteome analysis of proteins digested by trypsin was performed by LC MS/MS spectrometry. It was revealed that the ToEVs of band B and C contained 294 and 426 proteins, respectively. According to the sub-cellular localization predictions by PSORTb v3, ToEVs proteins consist of cytoplasmic protein, cytoplasmic membrane protein, cell wall protein, extracellular protein and various unknown proteins of the parental cell. Ratio of membrane protein to total protein in bands B and C ToEVs was slightly higher than that of parental cells. The analysis of predicted metabolic function of proteins from two major bands based on the COG (http://www.ncbi.nlm.nih.gov/COG/) database indicated that there was no big discrepancy between them. Proteomic comparison among parental cell, band B and band C ToEVs using Venn diagram showed ove ultracentrifugation. Two major bands (B & C) were obtained and theirs proteomes were further analyzed. ToEVs were spherical in shape and the size varied from 150 nm to 250 nm in diameter, and the vesicles of both band showed identical shape and size. The proteins of these two kinds of ToEVs were separated and identified by one-dimensional SDS-PAGE and the quantitative proteome analysis of proteins digested by trypsin was performed by LC MS/MS spectrometry. It was revealed that the ToEVs of band B and C contained 294 and 426 proteins, respectively. According to the sub-cellular localization predictions by PSORTb v3, ToEVs proteins consist of cytoplasmic protein, cytoplasmic membrane protein, cell wall protein, extracellular protein and various unknown proteins of the parental cell. Ratio of membrane protein to total protein in bands B and C ToEVs was slightly higher than that of parental cells. The analysis of predicted metabolic function of proteins from two major bands based on the COG (http://www.ncbi.nlm.nih.gov/COG/) database indicated that there was no big discrepancy between them. Proteomic comparison among parental cell, band B and band C ToEVs using Venn diagram showed ove
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/26293
Bibliographic Citation
Asia-Pacific Marine Biotechnology Conference, pp.pp115, 2014
Publisher
APMBC
Type
Conference
Language
English
Publisher
APMBC
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