4WMG

Structure of hen egg-white lysozyme from a microfludic harvesting device using synchrotron radiation (2.5A)

4WMG の概要

• エントリーDOI: 10.2210/pdb4wmg/pdb
• 分子名称: Lysozyme C (2 entities in total)
• 機能のキーワード: lysozyme, microfluidics, microcrystal, xfel, hydrolase
• 由来する生物種: Gallus gallus (Chicken)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 14331.16

構造登録者

Lyubimov, A.Y.,Murray, T.D.,Koehl, A.,Uervirojnangkoorn, M.,Zeldin, O.B.,Cohen, A.E.,Soltis, S.M.,Baxter, E.M.,Brewster, A.S.,Sauter, N.K.,Brunger, A.T.,Berger, J.M. (登録日: 2014-10-08, 公開日: 2015-04-22, 最終更新日: 2024-11-13)

主引用文献

Lyubimov, A.Y.,Murray, T.D.,Koehl, A.,Araci, I.E.,Uervirojnangkoorn, M.,Zeldin, O.B.,Cohen, A.E.,Soltis, S.M.,Baxter, E.L.,Brewster, A.S.,Sauter, N.K.,Brunger, A.T.,Berger, J.M.
Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array.
Acta Crystallogr.,Sect.D, 71:928-940, 2015

PubMed Abstract: X-ray free-electron lasers (XFELs) promise to enable the collection of interpretable diffraction data from samples that are refractory to data collection at synchrotron sources. At present, however, more efficient sample-delivery methods that minimize the consumption of microcrystalline material are needed to allow the application of XFEL sources to a wide range of challenging structural targets of biological importance. Here, a microfluidic chip is presented in which microcrystals can be captured at fixed, addressable points in a trap array from a small volume (<10 µl) of a pre-existing slurry grown off-chip. The device can be mounted on a standard goniostat for conducting diffraction experiments at room temperature without the need for flash-cooling. Proof-of-principle tests with a model system (hen egg-white lysozyme) demonstrated the high efficiency of the microfluidic approach for crystal harvesting, permitting the collection of sufficient data from only 265 single-crystal still images to permit determination and refinement of the structure of the protein. This work shows that microfluidic capture devices can be readily used to facilitate data collection from protein microcrystals grown in traditional laboratory formats, enabling analysis when cryopreservation is problematic or when only small numbers of crystals are available. Such microfluidic capture devices may also be useful for data collection at synchrotron sources.

PubMed: 25849403
DOI: 10.1107/S1399004715002308

実験手法

X-RAY DIFFRACTION (2.5 Å)