title: Three-stage refolding/unfolding of the dual-color beta-subunit in R-phycocyanin from Polysiphonia urceolata.
abstract: The conformational changes during refolding and unfolding of the dual-color beta-subunit in R-phycocyanin (R-PC) were monitored by the spectra, fluorescence anisotropy, and FRET. It was observed that both of the refolding and unfolding of the beta-subunit would undergo a three-stage conformational change, but in a reverse order. During the refolding process, at the first stage, the configuration of the tetrapyrrole chromophores transformed from the cyclohelical to the extended one, suggested by the blue-shifted spectra. At the second stage, recovery of the hydrogen-bond and hydrophobic interaction network fixed the chromophore in a more rigid configuration, suggested by a linear increase in the total fluorescence yield. At the third stage, the increase of the FRET efficiency suggested a protein-framework movement that made the two chromophores closer or/and into a more parallel orientation. The fluorescence anisotropy further confirmed the three-stage model.
authors.affiliation: Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, PR China.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Ma
authors.forename: Jie
authors.initials: J
authors.lastname: Xie
authors.forename: Chunxi
authors.initials: C
authors.lastname: Zhang
authors.forename: Jingquan
authors.initials: J
authors.lastname: Zhao
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D010798:Phycocyanin / Q000737:chemistry* / Q000648:ultrastructure*; D011489:Protein Denaturation; D017510:Protein Folding; D021122:Protein Subunits; D000461:Rhodophyta / Q000378:metabolism*
chemical_list: D021122:Protein Subunits; D010798:Phycocyanin