Effect of centrifugation on tryptic protein digestion

Soohwan Kim; Yeoseon Kim; Dabin Lee; Inyoung Kim; Jihyun Paek; Dongwon Shin; Jeongkwon Kim

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자료유형: 학술저널

저자정보: Soohwan Kim (Chungnam National University) Yeoseon Kim (Chungnam National University) Dabin Lee (Chungnam National University) Inyoung Kim (Chungnam National University) Jihyun Paek (Chungnam National University) Dongwon Shin (Chungnam National University) Jeongkwon Kim (Chungnam National University)

저널정보: 한국분석과학회 분석과학 분석과학 제30권 제2호

발행연도: 2017.4

수록면: 96 - 101 (6page)

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This study investigated the effect of centrifugation on tryptic digestion. This was done by applying different centrifugation speeds (6,000, 8,000, 10,000, 20,000, and 30,000×g) over various durations (0, 10, 20, 30, 40, 50, and 60 min) to digest two model proteins - cytochrome c and myoglobin. The intact proteins and resulting peptides were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Centrifugation greatly improved the tryptic digestion efficiency of cytochrome c, where either an increase in centrifugation speed or in digestion duration significantly improved the digestion of cytochrome c. However, centrifugation did not noticeably improve the digestion of myoglobin; 16 h of centrifuge-assisted tryptic digestion at 30,000×g barely removed the myoglobin protein peak. Similar results were also obtained when using conventional tryptic digestion with gentle mixing. When acetonitrile (ACN) was added to make 10% ACN buffer solutions, the myoglobin protein peak disappeared after 6 h of digestion using both centrifuge-assisted and conventional tryptic digestions.

#centrifuge #centrifugation #trypsin #digestion #proteins #MALDI

참고문헌 (7)

참고문헌 신청

W. Y. Chen and Y. C. Chen, Anal. Chem., 79(6), 2394- 2401 (2007). W. Sun, S. Gao, L. Wang, Y. Chen, S. Wu, X. Wang, D. Zheng, and Y. Gao, Mol. Cell. Proteomics, 5(10), S2-S2 (2006). D. Lopez-Ferrer, J. L. Capelo, and J. Vazquez, J. Proteome Res., 4(5), 1569-1574 (2005). H.-J. Yang, S. Shin, J. Kim, J. Hong, S. Lee, and J. Kim, Rapid Commun. Mass Spectrom., 25(1), 88-92 (2011). H. J. Yang, J. Hong, S. Lee, S. Shin, and J. Kim, Rapid Commun. Mass Spectrom., 24(7), 901-908 (2010).

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