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

FEBS Letters

Lavrik, O.I.²  Vavilin, V.I.¹  Maligin, E.G.¹  Tuzikov, F.V.¹  Ankilova, V.N.²  Moor, N.A.²  Zinoviev, V.V.¹ 

[1] All-Union Research Institute of Molecular Biology, Koltzovo, 633159 Novosibirsk Region, USSR;Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the USSR Academy of Sciences, 630090 Novosibirsk, USSR

关键词: Small-angle X-ray scattering;    Phenylalanyl-tRNA synthetase;    Enzyme-substrate interaction;    tRNAPhe;   

DOI  :  10.1016/0014-5793(88)80396-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 tRNA^Phe with phenylalanyl-tRNA synthetase have been determined and demonstrate negative cooperativity for binding of the two tRNA^Phe molecules. The structural parameters (R _g, R _c, semi-axes) have been determined for free phenylalanyl-tRNA synthetase and tRNA^Phe from E. coli MRE-600 and of enzyme complexes possessing one and two tRNA^Phe molecules, indicating structural rearrangements of the enzyme in the interaction with tRNA^Phe.

【 授权许可】

Unknown