1rnr
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(New page: 200px<br /><applet load="1rnr" size="450" color="white" frame="true" align="right" spinBox="true" caption="1rnr, resolution 2.5Å" /> '''AUTOCATALYTIC GENERAT...)
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AUTOCATALYTIC GENERATION OF DOPA IN THE ENGINEERED PROTEIN R2 F208Y FROM ESCHERICHIA COLI RIBONUCLEOTIDE REDUCTASE AND CRYSTAL STRUCTURE OF THE DOPA-208 PROTEIN
Overview
The mutant form Phe-208-->Tyr of the R2 protein of Escherichia coli, ribonucleotide reductase contains an intrinsic ferric-Dopa cofactor with, characteristic absorption bands at 460 and ca. 700 nm [Ormo, M., de Mare, F., Regnstrom, K., Aberg, A., Sahlin, M., Ling, J., Loehr, T. M., Sanders-Loehr, J., & Sjoberg, B. M. (1992) J. Biol. Chem. 267, 8711-8714]., The three-dimensional structure of the mutant protein, solved to 2.5-A, resolution, shows that the Dopa is localized to residue 208 and that it is, a bidentate ligand of Fe1 of the binuclear iron center of protein R2., Nascent apoR2 F208Y, lacking metal ions, can be purified from, overproducing cells grown in iron-depleted medium. ApoR2 F208Y is rapidly, and quantitatively converted to the Dopa-208 form in vitro by addition of, ferrous iron in the presence of oxygen. Other metal ions (Cu2+, Mn2+, Co2+) known to bind to the metal site of wild-type apoR2 do not generate a, Dopa in apoR2 F208Y. The autocatalytic generation of Dopa does not require, the presence of a tyrosine residue at position 122, the tyrosine which in, a wild-type R2 protein acquires the catalytically essential tyrosyl, radical. It is proposed that generation of Dopa initially follows the, suggested reaction mechanism for tyrosyl radical generation in the, wild-type protein and involves a ferryl intermediate, which in the case of, the mutant R2 protein oxygenates Tyr 208. This autocatalytic, metal-mediated reaction in the engineered R2 F208Y protein may serve as a, model for formation of covalently bound quinones in other proteins.
About this Structure
1RNR is a Single protein structure of sequence from Escherichia coli with FE and HG as ligands. Active as Ribonucleoside-diphosphate reductase, with EC number 1.17.4.1 Full crystallographic information is available from OCA.
Reference
Autocatalytic generation of dopa in the engineered protein R2 F208Y from Escherichia coli ribonucleotide reductase and crystal structure of the dopa-208 protein., Aberg A, Ormo M, Nordlund P, Sjoberg BM, Biochemistry. 1993 Sep 21;32(37):9845-50. PMID:8373782
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