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Science 3 March 2000: RESEARCH ARTICLE Vol. 287 no. 5458 pp. 1615-1622

Science 3 March 2000: RESEARCH ARTICLE Vol. 287 no. 5458 pp. 1615-1622 The Catalytic Pathway of Cytochrome P450cam at Atomic Resolution

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Science 3 March 2000: RESEARCH ARTICLE Vol. 287 no. 5458 pp. 1615-1622

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  1. Science 3 March 2000: RESEARCH ARTICLE Vol. 287 no. 5458 pp. 1615-1622 The Catalytic Pathway of Cytochrome P450cam at Atomic Resolution IlmeSchlichting, Joel Berendzen, Kelvin Chu, Ann M. Stock, Shelley A. Maves, David E. Benson, Robert M. Sweet, Dagmar Ringe, Gregory A. Petsko, Stephen G. Sligar

  2. The Catalytic Pathway of Cytochrome P450cam

  3. Joel Berendzen, IlmeSchlichting, Kelvin Chu

  4. P450cam: Dioxygen Complex

  5. P450cam: Activated Oxygen Intermediate

  6. Cytochrome P450: The Movie

  7. Ciba Foundation Symposium 1983

  8. Ciba Foundation Symposium 1983

  9. The Paradox

  10. IPMDH: Mesophile and Thermophile Zavodszky et al., PNAS (1998)

  11. The Paradox Hypothesis: Thermophilic enzymes are too rigid at moderate temperatures to function properly; they only exhibit the proper range of dynamics at their normally elevated temperatures

  12. Peter Zavodszky, TairoOshima and GerlindWallon

  13. H/D exchangeat 25o C Svingor et al., J. Biol. Chem. (2001) 276, 28121-28125

  14. H/D exchangenear temperature optimum Svingor et al., J. Biol. Chem. (2001) 276, 28121-28125

  15. IPMDH B-factors vs residueE. coli S. typhimuriumT. thermophilus

  16. DHFR catalyzes the reduction of DHF to THF using NADPH Mark Wilson

  17. DHFR

  18. DHFR: Reaction, Structure and Conformations of Met20 Loop Hod, Radkiewicz and Brooks, PNAS 2003

  19. NMR studies show protein motion that is probably important for ligand binding and product release Peter Wright

  20. E. coli DHFR 4oC

  21. ProcNatlAcadSci U S A. 108(39): 16247-16252 (2011)

  22. Fraser et al., PNAS 108(39): 16247-16252 (2011) We found that crystal cryocooling remodels the conformational distributions of more than 35% of side chains and eliminates packing defects necessary for functional motions. In the signaling switch protein, H-Ras, an allosteric network consistent with fluctuations detected in solution by NMR was uncovered in the room-temperature, but not the cryogenic, electron-density maps. These results expose a bias in structural databases toward smaller, overpacked, and unrealistically unique models. Monitoring room-temperature conformational ensembles by X-ray crystallography can reveal motions crucial for catalysis, ligand binding, and allosteric regulation.

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