title: The molecular interaction of a protein in highly concentrated solution investigated by Raman spectroscopy.
abstract: We used Raman spectroscopy to investigate the structure and interactions of lysozyme molecules in solution over a wide range of concentrations (2.5-300 mg ml(-1)). No changes in the amide-I band were observed as the concentration was increased, but the width of the Trp band at 1555 cm(-1) and the ratios of the intensities of the Tyr bands at 856 and 837 cm(-1), the Trp bands at 870 and 877 cm(-1), and the bands at 2940 (CH stretching) and 3420 cm(-1) (OH stretching) changed as the concentration was changed. These results reveal that although the distance between lysozyme molecules changed by more than an order of magnitude over the tested concentration range, the secondary structure of the protein did not change. The changes in the molecular interactions occurred in a stepwise process as the order of magnitude of the distance between molecules changed. These results suggest that Raman bands can be used as markers to investigate the behavior of high-concentration solutions of proteins and that the use of Raman spectroscopy will lead to progress in our understanding not only of the basic science of protein behavior under concentrated (i.e., crowded) conditions but also of practical processes involving proteins, such as in the field of biopharmaceuticals.
authors.affiliation: Department of Scientific & Semiconductor Instruments R&D, Application Development Center, Horiba Ltd., Minami-ku, Kyoto 601-8510, Japan.
authors.forename: Chikashi
authors.initials: C
authors.lastname: Ota
authors.forename: Shintaro
authors.initials: S
authors.lastname: Noguchi
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsumoto
mesh_terms: D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D011506:Proteins; D012996:Solutions
keywords: Raman spectroscopy; highly concentrated protein solution; molecular interaction