The anaerobic ribonucleotide reductase from Escherichia coli. The small protein is an activating enzyme containing a [4fe-4s](2+) center.
For deoxyribonucleotide synthesis during anaerobic growth, Escherichia coli cells depend on an oxygen-sensitive class III ribonucleotide reductase. The enzyme system consists of two proteins: protein alpha, on which ribonucleotides bind and are reduced, and protein beta, of which the function is to introduce a catalytically essential glycyl radical on protein alpha. Protein beta can assemble one [4Fe-4S] center per polypeptide enjoying both the [4Fe-4S](2+) and [4Fe-4S](1+) redox state, as shown by iron and sulfide analysis, Mössbauer spectroscopy (delta = 0.43 mm.s(-1), DeltaE(Q) = 1.0 mm.s(-1), [4Fe-4S](2+)), and EPR spectroscopy (g = 2. 03 and 1.93, [4Fe-4S](1+)). This iron center is sensitive to oxygen and can decompose into stable [2Fe-2S](2+) centers during exposure to air. This degraded form is nevertheless active, albeit to a lesser extent because of the conversion of the cluster into [4Fe-4S] forms during the strongly reductive conditions of the assay. Furthermore, protein beta has the potential to activate several molecules of protein alpha, suggesting that protein beta is an activating enzyme rather than a component of an alpha(2)beta(2) complex as previously claimed.
Tamarit
J
J
Laboratoire de Chimie et Biochimie des Centres Rédox Biologiques, Commissariat à l'Energie Atomique (CEA)/Département de Biologie Moléculaire et Structurale/Chimie et Biochimie 1087 CNRS, Université Joseph Fourier, 17 rue des Martyrs.
Mulliez
E
E
Meier
C
C
Trautwein
A
A
Fontecave
M
M
D000693:Anaerobiosis; D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D005609:Free Radicals; D005998:Glycine; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000378:metabolism*; D012264:Ribonucleotide Reductases / Q000737:chemistry / Q000378:metabolism*; D013053:Spectrophotometry; D015204:Spectroscopy, Mossbauer
D005609:Free Radicals; D007506:Iron-Sulfur Proteins; D012264:Ribonucleotide Reductases; D005998:Glycine