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Proteomic Profiling of Emiliania huxleyi Using a Three-Dimensional Separation Method Combined with Tandem Mass Spectrometry

Authors
Yun, GoyeunPark, Jong-MoonDuong, Van-AnMok, Jeong-HunJeon, JonghoNam, OnyouLee, JoonwonJin, EonSeonLee, Hookeun
Issue Date
Jul-2020
Publisher
MDPI
Keywords
Emiliania huxleyi; gene ontology; photosynthesis; proteomic profiling; three-dimensional (3D-LC) separation
Citation
MOLECULES, v.25, no.13
Journal Title
MOLECULES
Volume
25
Number
13
URI
https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/77995
DOI
10.3390/molecules25133028
ISSN
1420-3049
Abstract
Emiliania huxleyi is one of the most abundant marine planktons, and it has a crucial feature in the carbon cycle. However, proteomic analyses of Emiliania huxleyi have not been done extensively. In this study, a three-dimensional liquid chromatography (3D-LC) system consisting of strong cation exchange, high- and low-pH reversed-phase liquid chromatography was established for in-depth proteomic profiling of Emiliania huxleyi. From tryptic proteome digest, 70 fractions were generated and analyzed using liquid chromatography-tandem mass spectrometry. In total, more than 84,000 unique peptides and 10,000 proteins groups were identified with a false discovery rate of ≤0.01. The physicochemical properties of the identified peptides were evaluated. Using ClueGO, approximately 700 gene ontology terms and 15 pathways were defined from the identified protein groups with p-value ≤0.05, covering a wide range of biological processes, cellular components, and molecular functions. Many biological processes associated with CO2 fixation, photosynthesis, biosynthesis, and metabolic process were identified. Various molecular functions relating to protein binding and enzyme activities were also found. The 3D-LC strategy is a powerful approach for comparative proteomic studies on Emiliania huxleyi to reveal changes in its protein level and related mechanism.
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