The button test: A small scale method using microdialysis cells for assessing protein solubility at concentrations suitable for NMR

被引:34
作者
Bagby, S
Tong, KI
Liu, DJ
Alattia, JR
Ikura, M
机构
[1] UNIV TORONTO,ONTARIO CANC INST,DIV MOL & STRUCT BIOL,TORONTO,ON M5G 2M9,CANADA
[2] UNIV TORONTO,DEPT MED BIOPHYS,TORONTO,ON M5G 2M9,CANADA
基金
英国医学研究理事会;
关键词
microdialysis cell; NMR sample preparation; protein solubility; solution condition screen; TFIIB;
D O I
10.1023/A:1018359305544
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A simple method has been developed for screening solution conditions to determine conditions under which a protein is soluble at the high concentrations typically used for NMR spectroscopy. The method employs microdialysis cells or 'buttons'. The low sample volume (5 mu l) required for each microdialysis button permits testing of a wide range of solution conditions and temperatures with high protein concentrations, using a small amount of protein. Following precipitation of several NMR samples of the C-terminal core domain of human TFIIB, the microdialysis button screen facilitated identification of conditions in which precipitation of the TFIIB core domain was eliminated. The microdialysis button method for screening solution conditions is generally applicable and has been used to permit rapid identification of suitable NMR sample solution conditions for proteins involved in transcription and cell adhesion.
引用
收藏
页码:279 / 282
页数:4
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