TY - JOUR
T1 - Protein-ligand complex structure from serial femtosecond crystallography using soaked thermolysin microcrystals and comparison with structures from synchrotron radiation
AU - Naitow, Hisashi
AU - Matsuura, Yoshinori
AU - Tono, Kensuke
AU - Joti, Yasumasa
AU - Kameshima, Takashi
AU - Hatsui, Takaki
AU - Yabashi, Makina
AU - Tanaka, Rie
AU - Tanaka, Tomoyuki
AU - Sugahara, Michihiro
AU - Kobayashi, Jun
AU - Nango, Eriko
AU - Iwata, So
AU - Kunishima, Naoki
N1 - Funding Information:
This work was partly supported by the X-ray Free Electron Laser Priority Strategy Program from the Ministry of Education, Sports, Culture, Science and Technology, Japan (MEXT), the Strategic Basic Research Program from the Japan Science and Technology Agency (JST) and the SACLA Industry - Academia Partnership Program from RIKEN SPring-8 Center.
Publisher Copyright:
© Naitow et al. 2017.
PY - 2017/8
Y1 - 2017/8
N2 - Serial femtosecond crystallography (SFX) with an X-ray free-electron laser is used for the structural determination of proteins from a large number of microcrystals at room temperature. To examine the feasibility of pharmaceutical applications of SFX, a ligand-soaking experiment using thermolysin microcrystals has been performed using SFX. The results were compared with those from a conventional experiment with synchrotron radiation (SR) at 100K. A protein-ligand complex structure was successfully obtained from an SFX experiment using microcrystals soaked with a small-molecule ligand; both oil-based and water-based crystal carriers gave essentially the same results. In a comparison of the SFX and SR structures, clear differences were observed in the unit-cell parameters, in the alternate conformation of side chains, in the degree of water coordination and in the ligand-binding mode.The applicability of the ligand-soaking method in serial femtosecond crystallography has been examined to examine the feasibility of pharmaceutical applications of X-ray free-electron lasers.
AB - Serial femtosecond crystallography (SFX) with an X-ray free-electron laser is used for the structural determination of proteins from a large number of microcrystals at room temperature. To examine the feasibility of pharmaceutical applications of SFX, a ligand-soaking experiment using thermolysin microcrystals has been performed using SFX. The results were compared with those from a conventional experiment with synchrotron radiation (SR) at 100K. A protein-ligand complex structure was successfully obtained from an SFX experiment using microcrystals soaked with a small-molecule ligand; both oil-based and water-based crystal carriers gave essentially the same results. In a comparison of the SFX and SR structures, clear differences were observed in the unit-cell parameters, in the alternate conformation of side chains, in the degree of water coordination and in the ligand-binding mode.The applicability of the ligand-soaking method in serial femtosecond crystallography has been examined to examine the feasibility of pharmaceutical applications of X-ray free-electron lasers.
KW - X-ray crystallography
KW - X-ray free-electron laser
KW - diffraction before destruction
KW - microcrystal
KW - structure-based drug design
KW - thermolysin
UR - http://www.scopus.com/inward/record.url?scp=85026828777&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85026828777&partnerID=8YFLogxK
U2 - 10.1107/S2059798317008919
DO - 10.1107/S2059798317008919
M3 - Article
C2 - 28777085
AN - SCOPUS:85026828777
VL - 73
SP - 702
EP - 709
JO - Acta Crystallographica Section D: Structural Biology
JF - Acta Crystallographica Section D: Structural Biology
SN - 0907-4449
IS - 8
ER -