日本語
 
User Manual Privacy Policy ポリシー/免責事項 連絡先
  詳細検索ブラウズ

アイテム詳細


公開

学術論文

ENDOR spectroscopy and DFT calculations: Evidence for the hydrogen-bond network within α2 in the PCET of E. coli ribonucleotide reductase.

MPS-Authors
/persons/resource/persons14788

Argirevic,  T.
Research Group of Electron Paramagnetic Resonance, MPI for biophysical chemistry, Max Planck Society;

/persons/resource/persons14834

Bennati,  M.
Research Group of Electron Paramagnetic Resonance, MPI for biophysical chemistry, Max Planck Society;

フルテキスト (公開)

1569441.pdf
(出版社版), 503KB

付随資料 (公開)

1569441_si_001.pdf
(付録資料), 2MB

引用

Argirevic, T., Riplinger, C., Stubbe, J., Neese, F., & Bennati, M. (2012). ENDOR spectroscopy and DFT calculations: Evidence for the hydrogen-bond network within α2 in the PCET of E. coli ribonucleotide reductase. Journal of the American Chemical Society, 134(42), 17661-17670. doi:10.1021/ja3071682.


引用: http://hdl.handle.net/11858/00-001M-0000-0010-21CF-F
要旨
Escherichia coli class I ribonucleotide reductase (RNR) catalyzes the conversion of nucleotides to deoxynucleotides and is composed of two subunits: α2 and β2. β2 contains a stable di-iron tyrosyl radical (Y122•) cofactor required to generate a thiyl radical (C439•) in α2 over a distance of 35 Å, which in turn initiates the chemistry of the reduction process. The radical transfer process is proposed to occur by proton-coupled electron transfer (PCET) via a specific pathway: Y122 W48[?] Y356 in β2, across the subunit interface to Y731 Y730 C439 in α2. Within α2 a colinear PCET model has been proposed. To obtain evidence for this model, 3-amino tyrosine (NH2Y) replaced Y730 in α2, and this mutant was incubated with β2, cytidine 5′-diphosphate, and adenosine 5′-triphosphate to generate a NH2Y730• in D2O. [2H]-Electron–nuclear double resonance (ENDOR) spectra at 94 GHz of this intermediate were obtained, and together with DFT models of α2 and quantum chemical calculations allowed assignment of the prominent ENDOR features to two hydrogen bonds likely associated with C439 and Y731. A third proton was assigned to a water molecule in close proximity (2.2 Å O–H···O distance) to residue 730. The calculations also suggest that the unusual g-values measured for NH2Y730• are consistent with the combined effect of the hydrogen bonds to Cys439 and Tyr731, both nearly perpendicular to the ring plane of NH2Y730. The results provide the first experimental evidence for the hydrogen-bond network between the pathway residues in α2 of the active RNR complex, for which no structural data are available.