| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1987: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Research Abstract |
1.isolation and purification of cap binding protein,and characteristics of its amino acid composition
By preparing a Sepharose affinity column in which m^7GTP was covalently bound,the isolation and purification of the cap binding protein(CBP),which specifically binds with the cap structure of mRNA,were attempted from the rabbit reticulocyte ribosome. Since the protein was relatively unstable in usual physiological condition,the detailed physicochemical study on this protein is not yet complete at present. But,the preliminary analysis of its amino acid composition showed that CBP consists of the relatively large amount of tryptophan residue(4-6 residues), reflecting the characteristic of this protein.
2.Physicochemical studies on the interactions between aromatic amino caids and cap structure
The above-mentioned results imply that the aromatic amino acids such as tryptophan in CBP are important for the specific recognition of cap structure of mRNA. Therefore,the interactions between the ar
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omatic amino acids and m^7G analouges(as the model compound of cap structure) have been investigated. Spectroscopic and 1H-NMR studies in solution state showed that the order +f interaction force with m^7G was Trp>Tyr>Phe. Furtheremore,the temperature and concentration dependences of these amino acids for the interaction with m^7G gave the detailed informations as to the association constants and thermodynamical parameters. On the other hand,X-ray structural analyses provided the stacking parameters as to the aromatic ring-guanine base interaction at the atomic level.
3.Chemical synthesis of cap terminal structure and the interaction study with tryptophan-containing peptides
m^7GpppA, a terminal part of mRNA cap structure,was chemically stnthesized. The NMR studies for the interactions with a series of tryptophan-containing peptides showed that the stacking interaction between Trp indole ring and m^7G base and the hydrogen bonds between the carboxy1 group of acidic amino acid and the m^7G base are very important for the selective recognition of CBP with cap structure of mRNA. Less
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