title: Application of the small-angle X-ray scattering technique for the study of equilibrium enzyme-substrate interactions of phenylalanyl-tRNA synthetase from E. coli with tRNAPhe.
abstract: The small-angle X-ray scattering technique (SAXS) is proposed for the investigation of equilibrium macromolecular interactions of the enzyme-substrate type in solution. Experimental procedures and methods of analysing the data obtained from SAXS have been elaborated. The algorithm for the data analysis allows one to determine the stoichiometric, equilibrium and structural parameters of the enzyme-substrate complexes obtained. The thermodynamic characteristics for the formation of complexes of tRNAPhe with phenylalanyl-tRNA synthetase have been determined and demonstrate negative cooperativity for binding of the two tRNAPhe molecules. The structural parameters (Rg, Rc, semi-axes) have been determined for free phenylalanyl-tRNA synthetase and tRNAPhe from E. coli MRE-600 and of enzyme complexes possessing one and two tRNAPhe molecules, indicating structural rearrangements of the enzyme in the interaction with tRNAPhe.
authors.affiliation: All-Union Research Institute of Molecular Biology, Koltzovo, USSR.
authors.forename: F V
authors.initials: FV
authors.lastname: Tuzikov
authors.forename: V V
authors.initials: VV
authors.lastname: Zinoviev
authors.forename: V I
authors.initials: VI
authors.lastname: Vavilin
authors.forename: E G
authors.initials: EG
authors.lastname: Maligin
authors.forename: V N
authors.initials: VN
authors.lastname: Ankilova
authors.forename: N A
authors.initials: NA
authors.lastname: Moor
authors.forename: O I
authors.initials: OI
authors.lastname: Lavrik
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D010652:Phenylalanine-tRNA Ligase / Q000378:metabolism*; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000378:metabolism*; D012542:Scattering, Radiation; D012996:Solutions; D013816:Thermodynamics; D014965:X-Rays
chemical_list: D012345:RNA, Transfer, Amino Acid-Specific; D012360:RNA, Transfer, Phe; D012996:Solutions; D000604:Amino Acyl-tRNA Synthetases; D010652:Phenylalanine-tRNA Ligase