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Growth of large protein crystals by a large‐scale hanging‐drop method
Author(s) -
Kakinouchi Keisuke,
Nakamura Tsutomu,
Tamada Taro,
Adachi Hiroaki,
Sugiyama Shigeru,
Maruyama Mihoko,
Takahashi Yoshinori,
Takano Kazufumi,
Murakami Satoshi,
Inoue Tsuyoshi,
Kuroki Ryota,
Mori Yusuke,
Matsumura Hiroyoshi
Publication year - 2010
Publication title -
journal of applied crystallography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.429
H-Index - 162
ISSN - 1600-5767
DOI - 10.1107/s0021889810015967
Subject(s) - protein crystallization , drop (telecommunication) , mosaicity , crystal (programming language) , crystallization , materials science , crystallography , capillary action , ice crystals , diffraction , crystal growth , chemistry , optics , x ray crystallography , composite material , physics , telecommunications , organic chemistry , computer science , programming language
A method for growing large protein crystals is described. In this method, a cut pipette tip is used to hang large‐scale droplets (maximum volume 200 µl) consisting of protein and precipitating agents. A crystal grows at the vapor–liquid interface; thereafter the grown crystal can be retrieved by droplet–droplet contact both for repeated macroseeding and for mounting crystals in a capillary. Crystallization experiments with peroxiredoxin of Aeropyrum pernix K1 (thioredoxin peroxidase, ApTPx) and hen egg white lysozyme demonstrated that this large‐scale hanging‐drop method could produce a large‐volume crystal very effectively. A neutron diffraction experiment confirmed that an ApTPx crystal (6.2 mm 3 ) obtained by this method diffracted to beyond 3.5 Å resolution.