8SCY

Lysozyme crystallized in cyclic olefin copolymer-based microfluidic chips

8SCY の概要

• エントリーDOI: 10.2210/pdb8scy/pdb
• 分子名称: Lysozyme C (2 entities in total)
• 機能のキーワード: wild type, 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

Liu, Z.,Gu, K.,Shelby, M.L.,Gilbile, D.,Lyubimov, A.Y.,Russi, S.,Cohen, A.E.,Coleman, M.A.,Frank, M.,Kuhl, T.L. (登録日: 2023-04-05, 公開日: 2023-10-25, 最終更新日: 2024-10-23)

主引用文献

Liu, Z.,Gu, K.K.,Shelby, M.L.,Gilbile, D.,Lyubimov, A.Y.,Russi, S.,Cohen, A.E.,Narayanasamy, S.R.,Botha, S.,Kupitz, C.,Sierra, R.G.,Poitevin, F.,Gilardi, A.,Lisova, S.,Coleman, M.A.,Frank, M.,Kuhl, T.L.
A user-friendly plug-and-play cyclic olefin copolymer-based microfluidic chip for room-temperature, fixed-target serial crystallography.
Acta Crystallogr D Struct Biol, 79:944-952, 2023

PubMed Abstract: Over the past two decades, serial X-ray crystallography has enabled the structure determination of a wide range of proteins. With the advent of X-ray free-electron lasers (XFELs), ever-smaller crystals have yielded high-resolution diffraction and structure determination. A crucial need to continue advancement is the efficient delivery of fragile and micrometre-sized crystals to the X-ray beam intersection. This paper presents an improved design of an all-polymer microfluidic `chip' for room-temperature fixed-target serial crystallography that can be tailored to broadly meet the needs of users at either synchrotron or XFEL light sources. The chips are designed to be customized around different types of crystals and offer users a friendly, quick, convenient, ultra-low-cost and robust sample-delivery platform. Compared with the previous iteration of the chip [Gilbile et al. (2021), Lab Chip, 21, 4831-4845], the new design eliminates cleanroom fabrication. It has a larger imaging area to volume, while maintaining crystal hydration stability for both in situ crystallization or direct crystal slurry loading. Crystals of two model proteins, lysozyme and thaumatin, were used to validate the effectiveness of the design at both synchrotron (lysozyme and thaumatin) and XFEL (lysozyme only) facilities, yielding complete data sets with resolutions of 1.42, 1.48 and 1.70 Å, respectively. Overall, the improved chip design, ease of fabrication and high modifiability create a powerful, all-around sample-delivery tool that structural biologists can quickly adopt, especially in cases of limited sample volume and small, fragile crystals.

PubMed: 37747292
DOI: 10.1107/S2059798323007027

実験手法

X-RAY DIFFRACTION (1.45 Å)