Extended X-ray absorption fine structure studies on the iron-containing subunit of ribonucleotide reductase from Escherichia coli.
Iron K-edge X-ray absorption spectra were obtained on the protein B2, the small subunit of ribonucleotide reductase from Escherichia coli. Protein B2 contains a binuclear iron center with many properties in common with the iron center of oxidized hemerythrins. The extended X-ray absorption fine structure (EXAFS) measurements on protein B2 were analyzed and compared with published data for oxyhemerythrin. In protein B2 there are, in the first coordination shell around each Fe atom, five or six oxygen or nitrogen atoms that are directly coordinated ligands. In oxyhemerythrin there are six ligands to each iron. As in oxyhemerythrin, one of the ligands in the first shell of protein B2 is at a short distance, about 1.78 A, confirming the existence of a mu-oxo bridge. The other atoms of the first shell are at an average distance of 2.04 A, which is about 0.1 A shorter than in oxyhemerythrin. In protein B2 the Fe-Fe distance is in the range 3.26-3.48 A, and the bridging angle falls between 130 and 150 degrees. On the basis of these data, there is no direct evidence for any histidine ligands in protein B2, but the noise level leaves way for the possibility of a maximum of about three histidines for each Fe pair. The X-ray absorption spectrum of a hydroxyurea-treated sample was not significantly different from that of the native protein B2, which implies that no significant alteration in the structure of the iron site occurs upon destruction of the tyrosine radical.
Bunker
G
G
Institute for Structural and Functional Studies, Philadelphia, Pennsylvania 19104.
Petersson
L
L
Sjöberg
B M
BM
Sahlin
M
M
Chance
M
M
Chance
B
B
Ehrenberg
A
A
D004926:Escherichia coli / Q000201:enzymology*; D005583:Fourier Analysis; D006639:Histidine; D006918:Hydroxyurea; D007501:Iron / Q000032:analysis; D046911:Macromolecular Substances; D011485:Protein Binding; D012264:Ribonucleotide Reductases; D013057:Spectrum Analysis; D014965:X-Rays
D046911:Macromolecular Substances; D006639:Histidine; D007501:Iron; D012264:Ribonucleotide Reductases; D006918:Hydroxyurea