title: Structure of an E. coli integral membrane sulfurtransferase and its structural transition upon SCN(-) binding defined by EPR-based hybrid method.
abstract: Electron paramagnetic resonance (EPR)-based hybrid experimental and computational approaches were applied to determine the structure of a full-length E. coli integral membrane sulfurtransferase, dimeric YgaP, and its structural and dynamic changes upon ligand binding. The solution NMR structures of the YgaP transmembrane domain (TMD) and cytosolic catalytic rhodanese domain were reported recently, but the tertiary fold of full-length YgaP was not yet available. Here, systematic site-specific EPR analysis defined a helix-loop-helix secondary structure of the YagP-TMD monomers using mobility, accessibility and membrane immersion measurements. The tertiary folds of dimeric YgaP-TMD and full-length YgaP in detergent micelles were determined through inter- and intra-monomer distance mapping and rigid-body computation. Further EPR analysis demonstrated the tight packing of the two YgaP second transmembrane helices upon binding of the catalytic product SCN(-), which provides insight into the thiocyanate exportation mechanism of YgaP in the E. coli membrane.
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Shenglong
authors.initials: S
authors.lastname: Ling
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Yu
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Junhui
authors.initials: J
authors.lastname: Peng
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Cai
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xiong
authors.affiliation: National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL, 32310, U.S.A.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Hayati
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Longhua
authors.initials: L
authors.lastname: Zhang
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL, 32310, U.S.A.
authors.forename: Likai
authors.initials: L
authors.lastname: Song
authors.affiliation: National Laboratory for Physical Science at Microscale, School of Life Science, University of Science and Technology of China, and High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, 230027, P. R. China.
authors.forename: Changlin
authors.initials: C
authors.lastname: Tian
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008968:Molecular Conformation; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013466:Sulfurtransferases / Q000737:chemistry* / Q000378:metabolism; D013861:Thiocyanates / Q000737:chemistry* / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D013861:Thiocyanates; D013466:Sulfurtransferases; C031760:thiocyanate
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