Open AccessApproach for growth of high‐quality and large protein crystals
Author(s) -
Matsumura Hiroyoshi,
Sugiyama Shigeru,
Hirose Mika,
Kakinouchi Keisuke,
Maruyama Mihoko,
Murai Ryota,
Adachi Hiroaki,
Takano Kazufumi,
Murakami Satoshi,
Mori Yusuke,
Inoue Tsuyoshi
Publication year - 2011
Publication title -
journal of synchrotron radiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 99
ISSN - 1600-5775
DOI - 10.1107/s090904951003445x
Subject(s) - crystallization , agarose , protein crystallization , pipette , drop (telecommunication) , crystal (programming language) , chemistry , lysozyme , crystal growth , materials science , chromatography , chemical engineering , crystallography , computer science , biochemistry , telecommunications , organic chemistry , programming language , engineering
Three crystallization methods for growing large high‐quality protein crystals, i.e. crystallization in the presence of a semi‐solid agarose gel, top‐seeded solution growth (TSSG) and a large‐scale hanging‐drop method, have previously been presented. In this study the effectiveness of crystallization in the presence of a semi‐solid agarose gel has been further evaluated by crystallizing additional proteins in the presence of 2.0% ( w / v ) agarose gel, resulting in complete gelification with high mechanical strength. In TSSG the seed crystals are hung by a seed holder protruding from the top of the growth vessel to prevent polycrystallization. In the large‐scale hanging‐drop method, a cut pipette tip was used to maintain large‐scale droplets consisting of protein–precipitant solution. Here a novel crystallization method that combines TSSG and the large‐scale hanging‐drop method is reported. A large and single crystal of lysozyme was obtained by this method.