Structural and Thermodynamic Behavior of Eukaryotic Initiation Factor 4E in Supramolecular Formation with 4E-Binding Protein 1 and mRNA Cap Analogue, Studied by Spectroscopic Methods
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概要
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The structural and thermodynamic behavior of the complex formation of eIF4E with either or both mRNA cap analogue(m^7GTP, m^7GpppA, or m^7GpppG)and 4EBP1 has been investigated by spectroscopic measurements. Although the circular dichroism(CD)spectrum of eIF4E was little affected by the association with any cap analogue, the association constant of eIF4E with m^7GpppA/G, estimated from the fluorescence quenching, was about 10 times larger than that with m^7GTP. The van't Hoff analyses showed that the m^7GpppA/G binding is enthalpy-driven with a large negative ΔH^○, and this is in contrast with the entropy-driven binding of m^7GTP, where the positive ΔS^○ is large enough to overcome an increase of ΔH^○. This different behavior obviously originates in the interaction of the second nucleotide in m^7GpppA with eIF4E, suggesting the importance of the nucleotide sequence linked to the m^7Gppp terminal moiety, in addition to the specific interaction with the m^7G base, for the recognition of mRNA cap structure by eIF4E. On the other hand, the CD spectra indicated that the binding of 4EBP1, an endogenous eIF4E-regulatory protein without having any defined secondary structure, shifted the m^7GTP- or m^7GpppA/G-bound eIF4E to an irregular structure, although such a structural change was not observed for eIF4E alone. The association constant of 4EBP1 with m^7GTP- or m^7GpppA/G-bound eIF4E was by two orders of magnitude larger than that with eIF4E alone. These results suggest the close interrelation in the supramolecular formation of 4EBP-eIF4E-mRNA cap structure.
- 2001-10-01
著者
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Ishida Toshimasa
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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Tomoo Koji
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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Shen Xu
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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Uchiyama S
Faculty Of Pharmaceutical Sciences Osaka University
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Shen Xu
Department Of Chemistry Tongji University
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Uchiyama Susumu
Faculty Of Engineering Nagoya University
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Kobayashi Yuji
Faculty Of Education Tokushima University
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Kobayashi Yuji
Faculty Of Pharmaceutical Sciences Osaka University
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