spectroscopic studies of the iron and manganese reconstituted tyrosyl radical in bacillus cereus ribonucleotide reductase r2 protein光谱研究的铁和锰还原酪氨酰激进的蜡样芽胞杆菌r2核苷酸还原酶蛋白质.pdfVIP

Spectroscopic Studies of the Iron and Manganese Reconstituted Tyrosyl Radical in Bacillus Cereus Ribonucleotide Reductase R2 Protein 1 1 2 3 1 Ane B. Tomter , Giorgio Zoppellaro , Caleb B. Bell III , Anne-Laure Barra , Niels H. Andersen , Edward I. 2 1 Solomon , K. Kristoffer Andersson * 1 Department of Molecular Biosciences, University of Oslo, Oslo, Norway, 2 Department of Chemistry, Stanford University, Stanford, California, United States of America, ´ 3 Laboratoire National des Champs Magnetiques Intenses, LNCMI-G, UPR 3228, CNRS, Grenoble, France Abstract Ribonucleotide reductase (RNR) catalyzes the rate limiting step in DNA synthesis where ribonucleotides are reduced to the corresponding deoxyribonucleotides. Class Ib RNRs consist of two homodimeric subunits: R1E, which houses the active site; and R2F, which contains a metallo cofactor and a tyrosyl radical that initiates the ribonucleotide reduction reaction. We studied the R2F subunit of B. cereus reconstituted with iron or alternatively with manganese ions, then subsequently reacted with molecular oxygen to generate two tyrosyl-radicals. The two similar X-band EPR spectra did not change significantly over 4 to 50 K. From the 285 GHz EPR spectrum of the iron form, a g1-value of 2.0090 for the tyrosyl radical was extracted. This g1-value is similar to that observed in class Ia E. coli R2 and class Ib R2Fs with iron-oxygen cluster, suggesting the absence of hydrogen bond to the phenoxyl group. This was confirmed by resonance Raman spectroscopy, where the stretching vibration as