title: A method of cryoprotection for protein crystallography by using a microfluidic chip and its application for in situ X-ray diffraction measurements.
abstract: 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.
authors.affiliation: †Department of Molecular and Material Sciences, Interdisciplinary Graduate School of Engineering Sciences, Kyushu University, 6-1 Kasuga-Koen, Kasuga, Fukuoka 816-8580, Japan.|‡Division of Biotechnology and Macromolecular Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo 060-8628, Japan.|§Advanced Manufacturing Research Institute, National Institute of Advanced Industrial Science and Technology, 807-1 Shuku, Tosu, Saga 841-0052, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Maeki
authors.affiliation: ∥Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, United States.
authors.forename: Ashtamurthy S
authors.initials: AS
authors.lastname: Pawate
authors.affiliation: †Department of Molecular and Material Sciences, Interdisciplinary Graduate School of Engineering Sciences, Kyushu University, 6-1 Kasuga-Koen, Kasuga, Fukuoka 816-8580, Japan.|§Advanced Manufacturing Research Institute, National Institute of Advanced Industrial Science and Technology, 807-1 Shuku, Tosu, Saga 841-0052, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Yamashita
authors.affiliation: ⊥Kyushu Synchrotron Light Research Center, 8-7 Yayoigaoka, Tosu, Saga 841-0051, Japan.
authors.forename: Masahide
authors.initials: M
authors.lastname: Kawamoto
authors.affiliation: ‡Division of Biotechnology and Macromolecular Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo 060-8628, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Tokeshi
authors.affiliation: ∥Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, United States.
authors.forename: Paul J A
authors.initials: PJ
authors.lastname: Kenis
authors.affiliation: †Department of Molecular and Material Sciences, Interdisciplinary Graduate School of Engineering Sciences, Kyushu University, 6-1 Kasuga-Koen, Kasuga, Fukuoka 816-8580, Japan.|§Advanced Manufacturing Research Institute, National Institute of Advanced Industrial Science and Technology, 807-1 Shuku, Tosu, Saga 841-0052, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Miyazaki
mesh_terms: D000818:Animals; D002645:Chickens; D003461:Crystallography; D046210:Microfluidic Analytical Techniques; D009113:Muramidase / Q000737:chemistry* / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: C421864:hen egg lysozyme; D009113:Muramidase