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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2021.9
- 수록면
- 165 - 172 (8page)
- DOI
- 10.5010/JPB.2021.48.3.165
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초록· 키워드
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The solubilization of isolated proteins into the adequate buffer containing of surfactants is primary step for proteomic analysis. Particularly, sodium dodecyl sulfate (SDS) is the most widely used surfactant, however, it is not compatible with mass spectrometry (MS). Therefore, it must be removed prior to MS analysis through rigorous washing, which eventually results in inevitable loss. Recently, photocleavable surfactant, 4-hexylphenylazosulfonate (Azo), was reported which can be easily degraded by UV irradiation and is compatible with MS during proteomic approach using animal tissues. In this study, we employed comparative label-free proteomic analysis for evaluating the solubilization efficacy of the Azo and SDS surfactants using rice leave samples. This approach led to identification of 3,365 proteins and out of 682 proteins were determined as significantly modulated. Further, according to the subcellular localization prediction in SDS and Azo, proteins localized in the chloroplast were the major organelle accounting for 64% of the total organelle in the SDS sample, while only 37.5% of organelle proteins solubilized in the Azo were predicted to be localized in chloroplast. Taken together, this study validates the efficient solubilization of total protein isolated from plant material for bottom-up proteomics. Azo surfactant is suitable as substituents of SDS and promising for bottomup proteomics as it facilitates robust protein extraction, rapid washing step during enzymatic digestion, and MS analysis.
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참고문헌 (29)
참고문헌 신청
Alfonso-Garrido J / 2015 / Sample preparation strategies for improving the identification of membrane proteins by mass spectrometry / Anal Bioanal Chem 407:4893~4905
Bhuyan AK / 2010 / On the mechanism of SDS-induced protein denaturation / Biopolymers 93:186~199
Brown KA / 2019 / A photocleavable surfactant for top-down proteomics / Nat Methods 16:417~420
Brown KA / 2020 / High-throughput proteomics enabled by a photocleavable surfactant / Angew Chemie 132:8484~8488
Chang YH / 2015 / New massspectrometry-compatible degradable surfactant for tissue proteomics / J Proteome Res 14:1587~1599
Bhuyan AK / 2010 / On the mechanism of SDS-induced protein denaturation / Biopolymers 93:186~199
Brown KA / 2019 / A photocleavable surfactant for top-down proteomics / Nat Methods 16:417~420
Brown KA / 2020 / High-throughput proteomics enabled by a photocleavable surfactant / Angew Chemie 132:8484~8488
Chang YH / 2015 / New massspectrometry-compatible degradable surfactant for tissue proteomics / J Proteome Res 14:1587~1599
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