title: Picosecond time-resolved fluorescent spectroscopy of 1-anilino-8-naphthalene sulfonate binding with staphylococcal nuclease in the native and molten globule states.
abstract: We studied the picosecond time-resolved fluorescent spectroscopy of 1-anilino-8-naphthalene sulfonate (ANS), which binds to the staphylococcal nuclease (SNase) of the wild-type (WT) and the molten globule (MG) state. Three ANS emission bands at ∼530nm, ∼495nm, and ∼475nm are resolved, corresponding to three ANS states: the free ANS in solution and associated form adsorbing to surface sites and binding to active sites. The surface hydrophobicity of the WT is moderate and different from the MG state, as shown both in the position of the bands and by the concentration dependent ANS fluorescent decay. For MG, the decay of two blue bands accelerated with the increment of the ANS concentration, whereas the WT did not show this dependency. However, when pdTp, an inhibitor, was attached to the active site of the MG state, band 2 decay was also independent of the ANS concentration. These results indicate that the protein hydrophobic sites have two types of interactions with ANS.
authors.affiliation: Institute of Modern Optics, State Key Laboratory for Artificial Microstructure and Mesoscopic Physics, School of Physics, Peking University, Beijing 100871, China.
authors.forename: Guangyu
authors.initials: G
authors.lastname: Gao
authors.affiliation: Institute of Modern Optics, State Key Laboratory for Artificial Microstructure and Mesoscopic Physics, School of Physics, Peking University, Beijing 100871, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.affiliation: Institute of Modern Optics, State Key Laboratory for Artificial Microstructure and Mesoscopic Physics, School of Physics, Peking University, Beijing 100871, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Physics, Department of Chemistry and Biochemistry, and Programs of Biophysics, Chemical Physics and Biochemistry, The Ohio State University, Columbus, OH 43210, USA. Electronic address: zhong28@osu.edu.
authors.forename: Dongping
authors.initials: D
authors.lastname: Zhong
authors.affiliation: Institute of Modern Optics, State Key Laboratory for Artificial Microstructure and Mesoscopic Physics, School of Physics, Peking University, Beijing 100871, China. Electronic address: wangsf@pku.edu.cn.
authors.forename: Shufeng
authors.initials: S
authors.lastname: Wang
authors.affiliation: Institute of Modern Optics, State Key Laboratory for Artificial Microstructure and Mesoscopic Physics, School of Physics, Peking University, Beijing 100871, China.
authors.forename: Qihuang
authors.initials: Q
authors.lastname: Gong
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain; D057927:Hydrophobic and Hydrophilic Interactions; D008836:Micrococcal Nuclease / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence
chemical_list: D000817:Anilino Naphthalenesulfonates; D011994:Recombinant Proteins; D008836:Micrococcal Nuclease