title: High-throughput screening of optimal solution conditions for structural biological studies by fluorescence correlation spectroscopy.
abstract: 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.
authors.affiliation: Japan Biological Informatics Consortium (JBiC), Aomi, Koto-ku, Tokyo 135-8073, Japan.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Sugiki
authors.forename: Chie
authors.initials: C
authors.lastname: Yoshiura
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kofuku
authors.forename: Takumi
authors.initials: T
authors.lastname: Ueda
authors.forename: Ichio
authors.initials: I
authors.lastname: Shimada
authors.forename: Hideo
authors.initials: H
authors.lastname: Takahashi
mesh_terms: D054405:Chemokine CCL3 / Q000737:chemistry / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D054405:Chemokine CCL3; D046912:Multiprotein Complexes; D011506:Proteins; D017346:Protein Serine-Threonine Kinases