Observation of Asymmetry amongst Nucleotide Binding Sites of F_1-ATPase of Escherichia coli by ^(31)P NMR Spectroscopy

다국어 초록 (Multilingual Abstract)

It was regarded that the ^(31)P resonances of inherent nucleotides in F1-ATPase (EF1), as large as 380KDa, could not be observed by ^(31)P NMR spectroscopy. However, our ^(31)P NMR spectroscopy could differentiate between different nucleotide binding sites on EF1 from Escherichia coli. When EF1 was prepared in the absence of Mg^(2+), EF1 contained only ADP. Multiple ^(31)P resonances from β-phosphates of ADP bound to the EF1 were observed from the enzyme prepared without Mg^(2+), suggesting asymmetry or flexibility amongst nucleotide binding sites. ^(31)P resonances from enzyme bound ATP could be observed only from EF1, when the enzyme was prepared in the presence of Mg^(2+). This Mg^(2+) dependent ATP binding was very tight that, once bound, nucleotide could not be removed even after removal of Mg^(2+). ^(31)P NMR proved to be a valuable tool for investigating phosphorous related enzymes.

참고문헌 (Reference)

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