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Title: An isomorphous replacement method for efficient de novo phasing for serial femtosecond crystallography.
Authors: Yamashita, Keitaro
Pan, Dongqing  kyouindb  KAKEN_id  orcid (unconfirmed)
Okuda, Tomohiko
Sugahara, Michihiro
Kodan, Atsushi  kyouindb  KAKEN_id  orcid (unconfirmed)
Yamaguchi, Tomohiro  kyouindb  KAKEN_id
Murai, Tomohiro
Gomi, Keiko
Kajiyama, Naoki
Mizohata, Eiichi
Suzuki, Mamoru
Nango, Eriko  kyouindb  KAKEN_id
Tono, Kensuke
Joti, Yasumasa
Kameshima, Takashi
Park, Jaehyun
Song, Changyong
Hatsui, Takaki
Yabashi, Makina
Iwata, So  kyouindb  KAKEN_id
Kato, Hiroaki  kyouindb  KAKEN_id  orcid (unconfirmed)
Ago, Hideo
Yamamoto, Masaki
Nakatsu, Toru  kyouindb  KAKEN_id  orcid (unconfirmed)
Author's alias: 山下, 恵太郎
潘, 東青
村井, 智洋
登野, 健介
矢橋, 牧名
岩田, 想
加藤, 博章
吾郷, 日出夫
山本, 雅貴
中津, 亨
Keywords: Free-electron lasers
X-ray crystallography
Issue Date: 11-Sep-2015
Publisher: Nature Publishing Group
Journal title: Scientific reports
Volume: 5
Thesis number: 14017
Abstract: Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) holds great potential for structure determination of challenging proteins that are not amenable to producing large well diffracting crystals. Efficient de novo phasing methods are highly demanding and as such most SFX structures have been determined by molecular replacement methods. Here we employed single isomorphous replacement with anomalous scattering (SIRAS) for phasing and demonstrate successful application to SFX de novo phasing. Only about 20,000 patterns in total were needed for SIRAS phasing while single wavelength anomalous dispersion (SAD) phasing was unsuccessful with more than 80,000 patterns of derivative crystals. We employed high energy X-rays from SACLA (12.6 keV) to take advantage of the large anomalous enhancement near the LIII absorption edge of Hg, which is one of the most widely used heavy atoms for phasing in conventional protein crystallography. Hard XFEL is of benefit for de novo phasing in the use of routinely used heavy atoms and high resolution data collection.
Description: SACLAのX線自由電子レーザーを用いた新規タンパク質立体構造決定に世界で初めて成功. 京都大学プレスリリース. 2015-09-14.
Rights: This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit
DOI(Published Version): 10.1038/srep14017
PubMed ID: 26360462
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