5UVJ

Serial Millisecond Crystallography of Membrane and Soluble Protein Micro-crystals using Synchrotron Radiation

5UVJ の概要

• エントリーDOI: 10.2210/pdb5uvj/pdb
• 関連するPDBエントリー: 5UVI 5UVK 5UVL
• 分子名称: Lysozyme C, CHLORIDE ION, SODIUM ION, ... (4 entities in total)
• 機能のキーワード: 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: 14460.51

構造登録者

Martin-Garcia, J.M.,Conrad, C.E.,Nelson, G.,Stander, N.,Zatsepin, N.A.,Zook, J.,Zhu, L.,Geiger, J.,Chun, E.,Kissick, D.,Hilgart, M.C.,Ogata, C.,Ishchenko, A.,Nagaratnam, N.,Roy-Chowdhury, S.,Coe, J.,Subramanian, G.,Schaffer, A.,James, D.,Ketawala, G.,Venugopalan, N.,Xu, S.,Corcoran, S.,Ferguson, D.,Weierstall, U.,Spence, J.C.H.,Cherezov, V.,Fromme, P.,Fischetti, R.F.,Liu, W. (登録日: 2017-02-20, 公開日: 2017-05-24, 最終更新日: 2023-10-04)

主引用文献

Martin-Garcia, J.M.,Conrad, C.E.,Nelson, G.,Stander, N.,Zatsepin, N.A.,Zook, J.,Zhu, L.,Geiger, J.,Chun, E.,Kissick, D.,Hilgart, M.C.,Ogata, C.,Ishchenko, A.,Nagaratnam, N.,Roy-Chowdhury, S.,Coe, J.,Subramanian, G.,Schaffer, A.,James, D.,Ketwala, G.,Venugopalan, N.,Xu, S.,Corcoran, S.,Ferguson, D.,Weierstall, U.,Spence, J.C.H.,Cherezov, V.,Fromme, P.,Fischetti, R.F.,Liu, W.
Serial millisecond crystallography of membrane and soluble protein microcrystals using synchrotron radiation.
IUCrJ, 4:439-454, 2017

PubMed Abstract: Crystal structure determination of biological macromolecules using the novel technique of serial femtosecond crystallography (SFX) is severely limited by the scarcity of X-ray free-electron laser (XFEL) sources. However, recent and future upgrades render microfocus beamlines at synchrotron-radiation sources suitable for room-temperature serial crystallography data collection also. Owing to the longer exposure times that are needed at synchrotrons, serial data collection is termed serial millisecond crystallography (SMX). As a result, the number of SMX experiments is growing rapidly, with a dozen experiments reported so far. Here, the first high-viscosity injector-based SMX experiments carried out at a US synchrotron source, the Advanced Photon Source (APS), are reported. Microcrystals (5-20 µm) of a wide variety of proteins, including lysozyme, thaumatin, phycocyanin, the human A adenosine receptor (AAR), the soluble fragment of the membrane lipoprotein Flpp3 and proteinase K, were screened. Crystals suspended in lipidic cubic phase (LCP) or a high-molecular-weight poly(ethylene oxide) (PEO; molecular weight 8 000 000) were delivered to the beam using a high-viscosity injector. In-house data-reduction (hit-finding) software developed at APS as well as the SFX data-reduction and analysis software suites and enabled efficient on-site SMX data monitoring, reduction and processing. Complete data sets were collected for AAR, phycocyanin, Flpp3, proteinase K and lysozyme, and the structures of AAR, phycocyanin, proteinase K and lysozyme were determined at 3.2, 3.1, 2.65 and 2.05 Å resolution, respectively. The data demonstrate the feasibility of serial millisecond crystallography from 5-20 µm crystals using a high-viscosity injector at APS. The resolution of the crystal structures obtained in this study was dictated by the current flux density and crystal size, but upcoming developments in beamline optics and the planned APS-U upgrade will increase the intensity by two orders of magnitude. These developments will enable structure determination from smaller and/or weakly diffracting microcrystals.

PubMed: 28875031
DOI: 10.1107/S205225251700570X

実験手法

X-RAY DIFFRACTION (2.05 Å)