title: High-throughput protein refolding screening method using zeolite.
abstract: We established a 96-well-plate-based refolding screening system using zeolite. In this system, protein denatured and solubilized with 6 M guanidine hydrochloride is adsorbed onto zeolite placed in a 96-well plate. The refolding conditions can be tested by incubating the samples with refolding buffers under various conditions of pH, salts, and additives. In this study, we chose green fluorescent protein as the model protein. Green fluorescent protein was expressed as inclusion bodies, and we tested the effects of four pH conditions and six additives on its refolding. The results demonstrate that green fluorescent protein was more efficiently refolded with zeolite than with the conventional dilution method.
authors.affiliation: Research Center for Compact Chemical Process, AIST, Central 5, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
authors.forename: Takayuki Y
authors.initials: TY
authors.lastname: Nara
authors.forename: Hideaki
authors.initials: H
authors.lastname: Togashi
authors.forename: Chisato
authors.initials: C
authors.lastname: Sekikawa
authors.forename: Kengo
authors.initials: K
authors.lastname: Sakaguchi
authors.forename: Fujio
authors.initials: F
authors.lastname: Mizukami
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Tsunoda
mesh_terms: D001671:Biochemistry / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D019791:Guanidine / Q000737:chemistry; D011485:Protein Binding; D017510:Protein Folding; D017641:Zeolites / Q000737:chemistry*
chemical_list: D017641:Zeolites; D049452:Green Fluorescent Proteins; D019791:Guanidine