5GX4

Luciferin-regenerating enzyme collected with serial synchrotron rotational crystallography with accumulated dose of 14 MGy (12th measurement)

5GX4 の概要

• エントリーDOI: 10.2210/pdb5gx4/pdb
• 関連するPDBエントリー: 5GX1 5GX2 5GX3 5GX5
• 分子名称: Luciferin regenerating enzyme, MERCURY (II) ION, MAGNESIUM ION, ... (6 entities in total)
• 機能のキーワード: beta-prooeller, hydrolase
• 由来する生物種: Photinus pyralis (Common eastern firefly)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34955.63

構造登録者

Hasegawa, K.,Yamashita, K.,Murai, T.,Nuemket, N.,Hirata, K.,Ueno, G.,Ago, H.,Nakatsu, T.,Kumasaka, T.,Yamamoto, M. (登録日: 2016-09-15, 公開日: 2017-01-04, 最終更新日: 2023-11-08)

主引用文献

Hasegawa, K.,Yamashita, K.,Murai, T.,Nuemket, N.,Hirata, K.,Ueno, G.,Ago, H.,Nakatsu, T.,Kumasaka, T.,Yamamoto, M.
Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
J Synchrotron Radiat, 24:29-41, 2017

PubMed Abstract: Serial crystallography, in which single-shot diffraction images are collected, has great potential for protein microcrystallography. Although serial femtosecond crystallography (SFX) has been successfully demonstrated, limited beam time prevents its routine use. Inspired by SFX, serial synchrotron crystallography (SSX) has been investigated at synchrotron macromolecular crystallography beamlines. Unlike SFX, the longer exposure time of milliseconds to seconds commonly used in SSX causes radiation damage. However, in SSX, crystals can be rotated during the exposure, which can achieve efficient coverage of the reciprocal space. In this study, mercury single-wavelength anomalous diffraction (Hg-SAD) phasing of the luciferin regenerating enzyme (LRE) was performed using serial synchrotron rotation crystallography. The advantages of rotation and influence of dose on the data collected were evaluated. The results showed that sample rotation was effective for accurate data collection, and the optimum helical rotation step depended on multiple factors such as multiplicity and partiality of reflections, exposure time per rotation angle and the contribution from background scattering. For the LRE microcrystals, 0.25° was the best rotation step for the achievable resolution limit, whereas a rotation step larger than or equal to 1° was favorable for Hg-SAD phasing. Although an accumulated dose beyond 1.1 MGy caused specific damage at the Hg site, increases in resolution and anomalous signal were observed up to 3.4 MGy because of a higher signal-to-noise ratio.

PubMed: 28009544
DOI: 10.1107/S1600577516016362

実験手法

X-RAY DIFFRACTION (1.6 Å)