2,3-Difluorotyrosine at position 356 of ribonucleotide reductase R2: A probe of long-range proton-coupled electron transfer

Journal of the American Chemical Society

Volumn 125, Issue 35, 2003, Pages 10506-10507

  Yee, Cyril S ^a   Chang, Michelle C Y ^a   Ge, Jie ^a   Nocera, Daniel G ^a   Stubbe, JoAnne ^a  

^a MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

2,3 DIFLUOROTYROSINE; RIBONUCLEOTIDE REDUCTASE; TYROSINE DERIVATIVE; UNCLASSIFIED DRUG;

ARTICLE; COMPLEX FORMATION; DIFFERENTIAL PULSE VOLTAMMETRY; ELECTRON TRANSPORT; ENZYME ACTIVE SITE; OXIDATION; OXIDATION REDUCTION REACTION; PH;

AMINO ACID SEQUENCE; ELECTRONS; ESCHERICHIA COLI; HYDROGEN-ION CONCENTRATION; MODELS, MOLECULAR; MOLECULAR SEQUENCE DATA; OXIDATION-REDUCTION; PROTONS; RIBONUCLEOTIDE REDUCTASES; TYROSINE;

EID: 0041854730     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja036242r     Document Type: Article

References (24)

1. Jordan, A.; Reichard, P. Annu. Rev. Biochem. 1998, 67, 71-98.
2. Licht, S.; Stubbe, J. In Comprehensive Natural Products Chemistry; Barton, S. D., Nakanishi, K., Meth-Cohn, O., Poulter, C. D., Eds.; Elsevier: New York, 1999; Vol. 5, p 163.
3. Uhlin, U.; Eklund, H. Nature 1994, 370, 533-539.
4. -1 when donor and acceptor are in contact. See: Williams, R. J. B. J. Bioinorg. Chem. 1997, 2, 373-377.
5. (a) Stubbe, J.; Nocera, D. G.; Yee, C. S.; Chang, M. C. Y. Chem. Rev. 2003, 103, 2167-2201.
6. (b) Gray, H. B.; Winkler, J. R. In Electron Transfer in Chemistry; Balzani, V., Ed.; Wiley-VCH: Weinheim, Germany, 2001; Vol. 3. 1. 1, p 3.
7. (c) Page, C. C.; Moser, C. C.; Chen, X.; Dutton, P. L. Nature 1999, 402, 47-52.
8. (a) Ekberg, M.; Sahlin, M.; Eriksson, M.; Sjoberg, B. M. J. Biol. Chem. 1996, 271, 20655-20659.
9. (b) Ekberg, M.; Potsch, S.; Sandin, E.; Thunnissen, M.; Nordlund, P.; Sahlin, M.; Sjoberg, B. M. J. Biol. Chem. 1998, 273, 21003-21008.
10. (c) Rova, U.; Goodtzova, K.; Ingemarson, R.; Behravan, G.; Graslund, A.; Thelander, L. Biochemistry 1995, 34, 4267-4275.
11. (d) Rova, U.; Adrait, A.; Potsch, S.; Graslund, A.; Thelander, L. J. Biol. Chem. 1999, 274, 23746-23751.
12. (e) Siegbahn, P. E. M.; Eriksson, L.; Himo, F.; Pavlov, M. J. Phys. Chem. B 1998, 102, 10622-10629.
13. Tommos, C.; Skalicky, J. J.; Pilloud, D. L.; Wand, A. J.; Dutton, P. L. Biochemistry 1999, 38, 9495-9507.
14. Stubbe, J.; Van Der Donk, W. A. Chem. Rev. 1998, 98, 705-762.
15. Nordlund, P.; Eklund, H. J. Mol. Biol. 1993, 232, 123-164.
16. (a) Tong, W.; Burdi, D.; Riggs-Gelasco, P.; Chen, S.; Edmondson, D.; Huynh, B. H.; Stubbe, J.; Han, S.; Arvai, A.; Tainer, J. Biochemistry 1998, 37, 5840-5848.
17. (b) Climent, I.; Sjoberg, B. M.; Huang, C. Y. Biochemistry 1992, 31, 4801-4807.
18. -5 that of wt-R2 and is currently our lower limit of detection.
19. (a) Xu, M. Q.; Evans, T. C., Jr. Methods Enzymol. 2001, 24, 257-277.
20. (b) Ayers, B.; Blaschke, U.; Camarero, J.; Cotton, G. J.; Holford, M.; Muir, T. W. Biopolymers 1999, 51, 343-354.
21. Yee, C. S.; Seyedsayamdost, M. R.; Chang, M. C. Y.; Nocera, D. G.; Stubbe, J. Biochemistry 2003, submitted for publication.
22. From 10 L of LB-media, we obtain 20 g of cell paste, containing approximately 200 mg of the R2/intein/CBD construct. Given the 50% intein cleavage efficiency and the 70% ligation efficiency, we typically obtain ∼50 mg of purified full-length homodimeric protein. The detailed ligation and purification procedure will be found in ref 13.
23. Solid-phase peptide synthesis was performed with Fmoc protected amino acids and N-((dimethylamino)-1H-1,2,3-triazolo(4,5-b)pyridin-1-ylmethylene) -N-methylmethananaminium hexafluorophosphate N-oxide (HATU) coupling. Cysteine was protected with a tButhio, and residues D362 and S363 were replaced with a Fmoc-Asp(OtBu)-Ser(psiMe,Mepro)-OH dipeptide, which prevented extensive side reactions associated with cyclization and ring opening of aspartates. The synthesis was performed with standard procedures and will be found in ref 13.
24. Bollinger, J. M., Jr.; Tong, W. H.; Ravi, N.; Huynh, B. H.; Edmondson, D. E.; Stubbe, J. Methods Enzymol. 1995, 258, 278-303.