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High‐throughput screening of optimal solution conditions for structural biological studies by fluorescence correlation spectroscopy
Author(s) -
Sugiki Toshihiko,
Yoshiura Chie,
Kofuku Yutaka,
Ueda Takumi,
Shimada Ichio,
Takahashi Hideo
Publication year - 2009
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.92
Subject(s) - fluorescence correlation spectroscopy , fluorescence , fluorescence spectroscopy , protein aggregation , chemistry , biological system , characterization (materials science) , biophysics , molecule , nanotechnology , biochemistry , biology , materials science , physics , organic chemistry , quantum mechanics
Protein aggregation is an essential molecular event in a wide variety of biological situations, and is a causal factor in several degenerative diseases. The aggregation of proteins also frequently hampers structural biological analyses, such as solution NMR studies. Therefore, precise detection and characterization of protein aggregation are of crucial importance for various research fields. In this study, we demonstrate that fluorescence correlation spectroscopy (FCS) using a single‐molecule fluorescence detection system enables the detection of otherwise invisible aggregation of proteins at higher protein concentrations, which are suitable for structural biological experiments, and consumes relatively small amounts of protein over a short measurement time. Furthermore, utilizing FCS, we established a method for high‐throughput screening of protein aggregation and optimal solution conditions for structural biological experiments.