抄録
The hydrogen/deuterium exchange (HDX) method is useful to analyze kinetics of large macromolecular complexes, although its time resolution requires further improvement. A newly developed micro-reactor chip was made of polydimethylsiloxane with a 100-μm deep and wide microchannel. The channel in the chip has two mixing points of Y-shaped flow and allowed us to shorten time durations from the start to quenching for the HDX in 70S ribosome with high temporal resolution. This device enabled us to quench the deuterium incorporation at as early as 20 ms, detecting structural changes of individual ribosomal proteins in solution at the time scale comparable to a single reaction cycle for the peptide elongation. The profile of deuterium incorporation in individual proteins of the complex was superimposed on the X-ray crystal structure to depict the surface HDX map, revealing localization of protein movement in the ribosome. The current method serves as a useful method to visualize the regional movement of large macromolecules with high temporal resolution sufficient to examine protein dynamics.
| 元の言語 | English |
|---|---|
| ページ（範囲） | 132-138 |
| ページ数 | 7 |
| ジャーナル | International Journal of Mass Spectrometry |
| 巻 | 302 |
| 発行部数 | 1-3 |
| DOI | |
| 出版物ステータス | Published - 2011 4 30 |
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ASJC Scopus subject areas
- Physical and Theoretical Chemistry
- Spectroscopy
- Condensed Matter Physics
- Instrumentation
これを引用
Application of micro-reactor chip technique for millisecond quenching of deuterium incorporation into 70S ribosomal protein complex. / Yamamoto, Tatsuya; Shimizu, Yoshihiro; Ueda, Takuya; Shiro, Yoshitsugu; Suematsu, Makoto.
：: International Journal of Mass Spectrometry, 巻 302, 番号 1-3, 30.04.2011, p. 132-138.研究成果: Article
}
TY - JOUR
T1 - Application of micro-reactor chip technique for millisecond quenching of deuterium incorporation into 70S ribosomal protein complex
AU - Yamamoto, Tatsuya
AU - Shimizu, Yoshihiro
AU - Ueda, Takuya
AU - Shiro, Yoshitsugu
AU - Suematsu, Makoto
PY - 2011/4/30
Y1 - 2011/4/30
N2 - The hydrogen/deuterium exchange (HDX) method is useful to analyze kinetics of large macromolecular complexes, although its time resolution requires further improvement. A newly developed micro-reactor chip was made of polydimethylsiloxane with a 100-μm deep and wide microchannel. The channel in the chip has two mixing points of Y-shaped flow and allowed us to shorten time durations from the start to quenching for the HDX in 70S ribosome with high temporal resolution. This device enabled us to quench the deuterium incorporation at as early as 20 ms, detecting structural changes of individual ribosomal proteins in solution at the time scale comparable to a single reaction cycle for the peptide elongation. The profile of deuterium incorporation in individual proteins of the complex was superimposed on the X-ray crystal structure to depict the surface HDX map, revealing localization of protein movement in the ribosome. The current method serves as a useful method to visualize the regional movement of large macromolecules with high temporal resolution sufficient to examine protein dynamics.
AB - The hydrogen/deuterium exchange (HDX) method is useful to analyze kinetics of large macromolecular complexes, although its time resolution requires further improvement. A newly developed micro-reactor chip was made of polydimethylsiloxane with a 100-μm deep and wide microchannel. The channel in the chip has two mixing points of Y-shaped flow and allowed us to shorten time durations from the start to quenching for the HDX in 70S ribosome with high temporal resolution. This device enabled us to quench the deuterium incorporation at as early as 20 ms, detecting structural changes of individual ribosomal proteins in solution at the time scale comparable to a single reaction cycle for the peptide elongation. The profile of deuterium incorporation in individual proteins of the complex was superimposed on the X-ray crystal structure to depict the surface HDX map, revealing localization of protein movement in the ribosome. The current method serves as a useful method to visualize the regional movement of large macromolecules with high temporal resolution sufficient to examine protein dynamics.
KW - dynamics
KW - H/D exchange
KW - micro-reactor
KW - rapid quenching
KW - Ribosome
UR - http://www.scopus.com/inward/record.url?scp=79954630227&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79954630227&partnerID=8YFLogxK
U2 - 10.1016/j.ijms.2010.08.029
DO - 10.1016/j.ijms.2010.08.029
M3 - Article
AN - SCOPUS:79954630227
VL - 302
SP - 132
EP - 138
JO - International Journal of Mass Spectrometry
JF - International Journal of Mass Spectrometry
SN - 1387-3806
IS - 1-3
ER -