TY - JOUR
T1 - A method of cryoprotection for protein crystallography by using a microfluidic chip and its application for in situ X-ray diffraction measurements
AU - Maeki, Masatoshi
AU - Pawate, Ashtamurthy S.
AU - Yamashita, Kenichi
AU - Kawamoto, Masahide
AU - Tokeshi, Manabu
AU - Kenis, Paul J.A.
AU - Miyazaki, Masaya
N1 - Publisher Copyright:
© 2015 American Chemical Society.
PY - 2015/4/21
Y1 - 2015/4/21
N2 - We demonstrate a seamless and contactless method from protein crystallization to X-ray analysis using a microfluidic chip with the aim of obtaining a complete crystallographic data set of a protein crystal under cryogenic conditions. Our microfluidics-based approach did not require direct manipulation of the protein crystal. Therefore, the microfluidic chip approach is suitable for novices of X-ray analysis of protein crystals. We also investigated the effect of stepwise cryoprotection on the quality of protein crystals. Protein crystals with cryoprotection via on-chip manipulation did not show deterioration of crystallographic quality of the protein crystal. The complete diffraction data set of a protein crystal, which is required for determining the 3D structure of the target protein, is obtainable by a simple manipulation.
AB - We demonstrate a seamless and contactless method from protein crystallization to X-ray analysis using a microfluidic chip with the aim of obtaining a complete crystallographic data set of a protein crystal under cryogenic conditions. Our microfluidics-based approach did not require direct manipulation of the protein crystal. Therefore, the microfluidic chip approach is suitable for novices of X-ray analysis of protein crystals. We also investigated the effect of stepwise cryoprotection on the quality of protein crystals. Protein crystals with cryoprotection via on-chip manipulation did not show deterioration of crystallographic quality of the protein crystal. The complete diffraction data set of a protein crystal, which is required for determining the 3D structure of the target protein, is obtainable by a simple manipulation.
UR - http://www.scopus.com/inward/record.url?scp=84928338346&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84928338346&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.5b00151
DO - 10.1021/acs.analchem.5b00151
M3 - Article
C2 - 25834918
AN - SCOPUS:84928338346
SN - 0003-2700
VL - 87
SP - 4194
EP - 4200
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 8
ER -