title: Nondenaturing solubilization of beta2 microglobulin from inclusion bodies by L-arginine.
abstract: Expression of beta2 microglobulin (beta2m) in Escherichia coli resulted in formation of inclusion bodies. Attenuated total reflectance Fourier transform infrared analysis suggested a native-like secondary structure of beta2m in the inclusion bodies. Nondenaturing solubilization of the native-like beta2m from inclusion bodies was achieved using L-arginine solution, which enables an efficient recovery of beta2m with little aggregation. Greater beta2m solubilization from inclusion bodies was obtained at higher temperatures. Low-temperature solubilization yielded beta2m with fluorescence properties identical to those of native beta2m, but its secondary structure was slightly nonnative. Solubilization at moderate temperature gave beta2m with an apparently native structure. We propose an efficient nondenaturing solubilization method combining L-arginine and moderate temperature.
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, 6-6-07, Aoba-yama, Aoba-ku, Sendai 980-8579, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Umetsu
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsumoto
authors.forename: Shigeki
authors.initials: S
authors.lastname: Nitta
authors.forename: Tadafumi
authors.initials: T
authors.lastname: Adschiri
authors.forename: Daisuke
authors.initials: D
authors.lastname: Ejima
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Arakawa
authors.forename: Izumi
authors.initials: I
authors.lastname: Kumagai
mesh_terms: D001120:Arginine / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D002479:Inclusion Bodies / Q000737:chemistry*; D007202:Indicators and Reagents / Q000737:chemistry; D011489:Protein Denaturation; D012995:Solubility; D013696:Temperature; D001613:beta 2-Microglobulin / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D007202:Indicators and Reagents; D001613:beta 2-Microglobulin; D001120:Arginine