Oxidizing Intermediates in P450 Catalysis: A Case for Multiple Oxidants

  • Anuja R. Modi
  • John H. DawsonEmail author
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 851)


Cytochrome P450 (P450 or CYP) catalysis involves the oxygenation of organic compounds via a series of catalytic intermediates, namely, the ferric-peroxo, ferric-hydroperoxo, Compound I (Cpd I) and FeIII−(H2O2) intermediates. Now that the structures of P450 enzymes have been well established, a major focus of current research in the P450 area has been unraveling the intimate details and activities of these reactive intermediates. The general consensus is that the Cpd I intermediate is the most reactive species in the reaction cycle, especially when the reaction involves hydrocarbon hydroxylation. Cpd I has recently been characterized experimentally. Other than Cpd I, there is a multitude of evidence, both experimental as well as theoretical, supporting the involvement of other intermediates in various types of oxidation reactions. The involvement of these multiple oxidants has been experimentally demonstrated using P450 active-site mutants in epoxidation, heteroatom oxidation and dealkylation reactions. In this chapter, we will review the P450 reaction cycle and each of the reactive intermediates to discuss their role in oxidation reactions.


Cytochrome P450 Reaction cycle Compound I Ferric-peroxo Ferric-hydroperoxo Reactive intermediates Multiple oxidants 



The NIH (GM-26730) has supported cytochrome P450 research in the Dawson laboratory. We would like to thank Dr. Masanori Sono for pertinent advice.


  1. 1.
    Poulos TL, Johnson EF (2005) Structures of cytochrome P450 enzymes. In: Ortiz de Montellano PR (ed) Cytochrome P450: structure, mechanism, and biochemistry, 3rd edn. Kluwer Academic/Plenum Publishers, New York, pp 87–114CrossRefGoogle Scholar
  2. 2.
    Sono M, Roach MP, Coulter ED, Dawson JH (1996) Heme-containing oxygenases. Chem Rev 96:2841–2888CrossRefPubMedGoogle Scholar
  3. 3.
    Omura T, Sato R (1964) The carbon monoxide-binding pigment of liver microsomes: I. Evidence for its hemoprotein nature. J Biol Chem 239:2370–2378PubMedGoogle Scholar
  4. 4.
    Filatov M, Reckien W, Peyerimhoff SD, Shaik S (2000) What are the reasons for the kinetic stability of a mixture of H2 and O2? J Phys Chem A 104:12014–12020CrossRefGoogle Scholar
  5. 5.
    Blanksby SJ, Ellison GB (2003) Bond dissociation energies of organic molecules. Acc Chem Res 36:255–263CrossRefPubMedGoogle Scholar
  6. 6.
    Sligar SG, Makris TM, Denisov IG (2005) Thirty years of microbial P450 monooxygenase research: peroxo-heme intermediates—the central bus station in heme oxygenase catalysis. Biochem Biophys Res Commun 338:346–354CrossRefPubMedGoogle Scholar
  7. 7.
    Meunier B (1992) Metalloporphyrins as versatile catalysts for oxidation reactions and oxidative DNA cleavage. Chem Rev 92:1411–1456CrossRefGoogle Scholar
  8. 8.
    Glieder A, Farinas ET, Arnold FH (2002) Laboratory evolution of a soluble, self-sufficient, highly active alkane hydroxylase. Nat Biotechnol 20:1135–1139CrossRefPubMedGoogle Scholar
  9. 9.
    Schlichting I, Berendzen J, Chu K, Stock AM, Maves SA, Benson DE, Sweet RM, Ringe D, Petsko GA, Sligar SG (2000) The catalytic pathway of cytochrome P450cam at atomic resolution. Science 287:1615–1622CrossRefPubMedGoogle Scholar
  10. 10.
    Nebert DW, Gonzalez FJ (1987) P450 genes: structure, evolution, and regulation. Annu Rev Biochem 56:945–993CrossRefPubMedGoogle Scholar
  11. 11.
    Narhi LO, Fulco AJ (1986) Characterization of a catalytically self-sufficient 119,000-dalton cytochrome P-450 monooxygenase induced by barbiturates in Bacillus megaterium. J Biol Chem 261:7160–7169PubMedGoogle Scholar
  12. 12.
    Ruettinger RT, Wen LP, Fulco AJ (1989) Coding nucleotide, 5′ regulatory, and deduced amino acid sequences of P-450BM-3, a single peptide cytochrome P-450:NADPH-P-450 reductase from Bacillus megaterium. J Biol Chem 264:10987–10995PubMedGoogle Scholar
  13. 13.
    Poulos TL, Finzel BC, Gunsalus IC, Wagner GC, Kraut J (1985) The 2.6-Å crystal structure of Pseudomonas putida cytochrome P-450. J Biol Chem 260:16122–16130PubMedGoogle Scholar
  14. 14.
    Gunsalus IC, Pederson TC, Sligar SG (1975) Oxygenase-catalyzed biological hydroxylations. Annu Rev Biochem 44:377–407CrossRefPubMedGoogle Scholar
  15. 15.
    Meunier B, de Visser SP, Shaik S (2004) Mechanism of oxidation reactions catalyzed by cytochrome P450 enzymes. Chem Rev 104:3947–3980CrossRefPubMedGoogle Scholar
  16. 16.
    Winfield ME (1965) Mechanisms of oxygen uptake: the autoxidation of myoglobin and of reduced cyanocobaltates and their significance to oxidase reactions. In: King TE, Mason HS, Morrison M (eds) Oxidases and related redox systems, vol 1. Wiley, New York, pp 115–130Google Scholar
  17. 17.
    Dunford HB, Stillman JS (1976) On the function and mechanism of action of peroxidases. Coord Chem Rev 19:187–251CrossRefGoogle Scholar
  18. 18.
    Imai M, Shimada H, Watanabe Y, Matsushima-Hibiya Y, Makino R, Koga H, Horiuchi T, Ishimura Y (1989) Uncoupling of the cytochrome P-450cam monooxygenase reaction by a single mutation, threonine-252 to alanine or valine: possible role of the hydroxy amino acid in oxygen activation. Proc Natl Acad Sci U S A 86:7823–7827CrossRefPubMedCentralPubMedGoogle Scholar
  19. 19.
    Egawa T, Shimada H, Ishimura Y (1994) Evidence for compound I formation in the reaction of cytochrome P450cam with m-chloroperbenzoic acid. Biochem Biophys Res Commun 201:1464–1469CrossRefPubMedGoogle Scholar
  20. 20.
    Schünemann V, Lendzian F, Jung C, Contzen J, Barra AL, Sligar SG, Trautwein AX (2004) Tyrosine radical formation in the reaction of wild type and mutant cytochrome P450cam with peroxy acids: a multifrequency EPR study of intermediates on the millisecond time scale. J Biol Chem 279:10919–10930CrossRefPubMedGoogle Scholar
  21. 21.
    Spolitak T, Dawson JH, Ballou DP (2005) Reaction of ferric cytochrome P450cam with peracids: kinetic characterization of intermediates on the reaction pathway. J Biol Chem 280:20300–20309CrossRefPubMedGoogle Scholar
  22. 22.
    Shimada H, Watanabe Y, Imai M, Makino R, Koga H, Horiuchi T, Ishimura Y (1991) The role of threonine 252 in the oxygen activation by cytochrome P-450 cam: mechanistic studies by site-directed mutagenesis. In: Simandi LI (ed) Dioxygen activation and homogeneous catalytic oxidation. Elsevier, Amsterdam, pp 3136–3319Google Scholar
  23. 23.
    Auclair K, Moënne-Loccoz P, Ortiz de Montellano PR (2001) Roles of the proximal heme thiolate ligand in cytochrome P450cam. J Am Chem Soc 123:4877–4885CrossRefPubMedGoogle Scholar
  24. 24.
    Perera R, Sono M, Sigman JA, Pfister TD, Lu Y, Dawson JH (2003) Neutral thiol as a proximal ligand to ferrous heme iron: implications for heme proteins that lose cysteine thiolate ligation on reduction. Proc Natl Acad Sci U S A 100:3641–3646CrossRefPubMedCentralPubMedGoogle Scholar
  25. 25.
    Yoshioka S, Takahashi S, Ishimori K, Morishima I (2000) Roles of the axial push effect in cytochrome P450cam studied with the site-directed mutagenesis at the heme proximal site. J Inorg Biochem 81:141–151CrossRefPubMedGoogle Scholar
  26. 26.
    Yoshioka S, Tosha T, Takahashi S, Ishimori K, Hori H, Morishima I (2002) Roles of the proximal hydrogen bonding network in cytochrome P450cam-catalyzed oxygenation. J Am Chem Soc 124:14571–14579CrossRefPubMedGoogle Scholar
  27. 27.
    Dawson JH (1988) Probing structure-function relations in heme-containing oxygenases and peroxidases. Science 240:433–439CrossRefPubMedGoogle Scholar
  28. 28.
    Dawson JH, Holm RH, Trudell JR, Barth G, Linder RE, Bunnenberg E, Djerassi C, Tang SC (1976) Oxidized cytochrome P-450. Magnetic circular dichroism evidence for thiolate ligation in the substrate-bound form. Implications for the catalytic mechanism. J Am Chem Soc 98:3707–3709CrossRefPubMedGoogle Scholar
  29. 29.
    Poulos TL, Kraut J (1980) The stereochemistry of peroxidase catalysis. J Biol Chem 255:8199–8205PubMedGoogle Scholar
  30. 30.
    Martinis SA, Atkins WM, Stayton PS, Sligar SG (1989) A conserved residue of cytochrome P-450 is involved in heme-oxygen stability and activation. J Am Chem Soc 111:9252–9253CrossRefGoogle Scholar
  31. 31.
    Yeom H, Sligar SG, Li H, Poulos TL, Fulco AJ (1995) The role of Thr268 in oxygen activation of cytochrome P450BM-3. Biochemistry 34:14733–14740CrossRefPubMedGoogle Scholar
  32. 32.
    Imai Y, Nakamura M (1988) The importance of threonine-301 from cytochromes P-450 (laurate (ω-1)-hydroxylase and testosterone 16α-hydroxylase) in substrate binding as demonstrated by site-directed mutagenesis. FEBS Lett 234:313–315CrossRefPubMedGoogle Scholar
  33. 33.
    Raag R, Martinis SA, Sligar SG, Poulos TL (1991) Crystal structure of the cytochrome P-450CAM active site mutant Thr252Ala. Biochemistry 30:11420–11429CrossRefPubMedGoogle Scholar
  34. 34.
    Harris DL, Loew GH (1996) Investigation of the proton-assisted pathway to formation of the catalytically active, ferryl species of P450s by molecular dynamics studies of P450eryF. J Am Chem Soc 118:6377–6387CrossRefPubMedGoogle Scholar
  35. 35.
    Truan G, Peterson JA (1998) Thr268 in substrate binding and catalysis in P450BM-3. Arch Biochem Biophys 349:53–64CrossRefPubMedGoogle Scholar
  36. 36.
    Vidakovic M, Sligar SG, Li H, Poulos TL (1998) Understanding the role of the essential Asp251 in cytochrome P450cam using site-directed mutagenesis: crystallography, and kinetic solvent isotope effect. Biochemistry 37:9211–9219CrossRefPubMedGoogle Scholar
  37. 37.
    Gerber NC, Sligar SG (1994) A role for Asp-251 in cytochrome P-450cam oxygen activation. J Biol Chem 269:4260–4266PubMedGoogle Scholar
  38. 38.
    Rittle J, Green MT (2010) Cytochrome P450 compound I: capture, characterization, and C-H bond activation kinetics. Science 330:933–937CrossRefPubMedGoogle Scholar
  39. 39.
    Loida PJ, Sligar SG (1993) Molecular recognition in cytochrome P-450: mechanism for the control of uncoupling reactions. Biochemistry 32:11530–11538CrossRefPubMedGoogle Scholar
  40. 40.
    Eisenstein L, Debey P, Douzou P (1977) P450cam: oxygenated complexes stabilized at low temperature. Biochem Biophys Res Commun 77:1377–1383CrossRefPubMedGoogle Scholar
  41. 41.
    Antonini E, Brunori M (1971) Hemoglobin and myoglobin in their reactions with ligands. North-Holland, AmsterdamGoogle Scholar
  42. 42.
    Couture M, Stuehr DJ, Rousseau DL (2000) The ferrous dioxygen complex of the oxygenase domain of neuronal nitric-oxide synthase. J Biol Chem 275:3201–3205CrossRefPubMedGoogle Scholar
  43. 43.
    Macdonald IDG, Sligar SG, Christian JF, Unno M, Champion PM (1998) Identification of the Fe−O−O bending mode in oxycytochrome P450cam by resonance Raman spectroscopy. J Am Chem Soc 121:376–380CrossRefGoogle Scholar
  44. 44.
    Sono M, Eble KS, Dawson JH, Hager LP (1985) Preparation and properties of ferrous chloroperoxidase complexes with dioxygen, nitric oxide, and an alkyl isocyanide. Spectroscopic dissimilarities between the oxygenated forms of chloroperoxidase and cytochrome P-450. J Biol Chem 260:15530–15535PubMedGoogle Scholar
  45. 45.
    Bangcharoenpaurpong O, Rizos AK, Champion PM, Jollie D, Sligar SG (1986) Resonance Raman detection of bound dioxygen in cytochrome P-450cam. J Biol Chem 261:8089–8092PubMedGoogle Scholar
  46. 46.
    Fischer RT, Trzaskos JM, Magolda RL, Ko SS, Brosz CS, Larsen B (1991) Lanosterol 14α-methyl demethylase. Isolation and characterization of the third metabolically generated oxidative demethylation intermediate. J Biol Chem 266:6124–6132PubMedGoogle Scholar
  47. 47.
    Akhtar M, Corina D, Miller S, Shyadehi AZ, Wright JN (1994) Mechanism of the acyl-carbon cleavage and related reactions catalyzed by multifunctional P-450s: studies on cytochrome P-45017α. Biochemistry 33:4410–4418CrossRefPubMedGoogle Scholar
  48. 48.
    Akhtar M, Alexander K, Boar RB, McGhie JF, Barton DH (1978) Chemical and enzymic studies on the characterization of intermediates during the removal of the 14α-methyl group in cholesterol biosynthesis. The use of 32-functionalized lanostane derivatives. Biochem J 169:449–463PubMedCentralPubMedGoogle Scholar
  49. 49.
    Corina DL, Miller SL, Wright JN, Akhtar M (1991) The mechanism of cytochrome P-450 dependent C-C bond cleavage: studies on 17α-hydroxylase-17,20-lyase. J Chem Soc Chem Commun 782–783Google Scholar
  50. 50.
    Roberts ES, Vaz AD, Coon MJ (1991) Catalysis by cytochrome P-450 of an oxidative reaction in xenobiotic aldehyde metabolism: deformylation with olefin formation. Proc Natl Acad Sci U S A 88:8963–8966CrossRefPubMedCentralPubMedGoogle Scholar
  51. 51.
    Kuo CL, Raner GM, Vaz ADN, Coon MJ (1999) Discrete species of activated oxygen yield different cytochrome P450 heme adducts from aldehydes. Biochemistry 38:10511–10518CrossRefPubMedGoogle Scholar
  52. 52.
    Bestervelt LL, Vaz AD, Coon MJ (1995) Inactivation of ethanol-inducible cytochrome P450 and other microsomal P450 isozymes by trans-4-hydroxy-2-nonenal, a major product of membrane lipid peroxidation. Proc Natl Acad Sci U S A 92:3764–3768CrossRefPubMedCentralPubMedGoogle Scholar
  53. 53.
    Korth HG, Sustmann R, Thater C, Butler AR, Ingold KU (1994) On the mechanism of the nitric oxide synthase-catalyzed conversion of N ω-hydroxy-L-arginine to citrulline and nitric oxide. J Biol Chem 269:17776–17779PubMedGoogle Scholar
  54. 54.
    Wertz DL, Sisemore MF, Selke M, Driscoll J, Valentine JS (1998) Mimicking cytochrome P-450 2B4 and aromatase: aromatization of a substrate analogue by a peroxo Fe(III) porphyrin complex. J Am Chem Soc 120:5331–5332CrossRefGoogle Scholar
  55. 55.
    Sisemore MF, Burstyn JN, Valentine JS (1996) Epoxidation of electron-deficient olefins by a nucleophilic iron(III) peroxo porphyrinato complex, peroxo(tetramesitylporphyrinato)ferrate(1−). Angew Chem Int Ed 35:206–208CrossRefGoogle Scholar
  56. 56.
    Davydov R, Macdonald IDG, Makris TM, Sligar SG, Hoffman BM (1999) EPR and ENDOR of catalytic intermediates in cryoreduced native and mutant oxy-cytochromes P450cam: mutation-induced changes in the proton delivery system. J Am Chem Soc 121:10654–10655CrossRefGoogle Scholar
  57. 57.
    Davydov R, Makris TM, Kofman V, Werst DE, Sligar SG, Hoffman BM (2001) Hydroxylation of camphor by reduced oxy-cytochrome P450cam: mechanistic implications of EPR and ENDOR studies of catalytic intermediates in native and mutant enzymes. J Am Chem Soc 123:1403–1415CrossRefPubMedGoogle Scholar
  58. 58.
    Vaz ADN, McGinnity DF, Coon MJ (1998) Epoxidation of olefins by cytochrome P450: evidence from site-specific mutagenesis for hydroperoxo-iron as an electrophilic oxidant. Proc Natl Acad Sci U S A 95:3555–3560CrossRefPubMedCentralPubMedGoogle Scholar
  59. 59.
    Jin S, Makris TM, Bryson TA, Sligar SG, Dawson JH (2003) Epoxidation of olefins by hydroperoxo−ferric cytochrome P450. J Am Chem Soc 125:3406–3407CrossRefPubMedGoogle Scholar
  60. 60.
    de Visser SP, Ogliaro F, Harris N, Shaik S (2001) Multi-state epoxidation of ethene by cytochrome P450: a quantum chemical study. J Am Chem Soc 123:3037–3047CrossRefPubMedGoogle Scholar
  61. 61.
    de Visser SP, Ogliaro F, Sharma PK, Shaik S (2002) What factors affect the regioselectivity of oxidation by cytochrome P450? A DFT study of allylic hydroxylation and double bond epoxidation in a model reaction. J Am Chem Soc 124:11809–11826CrossRefPubMedGoogle Scholar
  62. 62.
    Ogliaro F, Cohen S, de Visser SP, Shaik S (2000) Medium polarization and hydrogen bonding effects on compound I of cytochrome P450: what kind of a radical is it really? J Am Chem Soc 122:12892–12893CrossRefGoogle Scholar
  63. 63.
    Ogliaro F, de Visser SP, Cohen S, Sharma PK, Shaik S (2002) Searching for the second oxidant in the catalytic cycle of cytochrome P450: a theoretical investigation of the iron(III)-hydroperoxo species and its epoxidation pathways. J Am Chem Soc 124:2806–2817CrossRefPubMedGoogle Scholar
  64. 64.
    de Visser SP, Ogliaro F, Sharma PK, Shaik S (2002) Hydrogen bonding modulates the selectivity of enzymatic oxidation by P450: chameleon oxidant behavior by compound I. Angew Chem Int Ed 41:1947–1951CrossRefGoogle Scholar
  65. 65.
    Volz TJ, Rock DA, Jones JP (2002) Evidence for two different active oxygen species in cytochrome P450 BM3 mediated sulfoxidation and N-dealkylation reactions. J Am Chem Soc 124:9724–9725CrossRefPubMedGoogle Scholar
  66. 66.
    Watanabe Y (2001) Alternatives to the oxoferryl porphyrin cation radical as the proposed reactive intermediate of cytochrome P450: two-electron oxidized Fe(III) porphyrin derivatives. J Biol Inorg Chem 6:846–856CrossRefPubMedGoogle Scholar
  67. 67.
    Newcomb M, Shen R, Choi SY, Toy PH, Hollenberg PF, Vaz ADN, Coon MJ (2000) Cytochrome P450-catalyzed hydroxylation of mechanistic probes that distinguish between radicals and cations. Evidence for cationic but not for radical intermediates. J Am Chem Soc 122:2677–2686CrossRefGoogle Scholar
  68. 68.
    Jin S, Bryson T, Dawson J (2004) Hydroperoxoferric heme intermediate as a second electrophilic oxidant in cytochrome P450-catalyzed reactions. J Biol Inorg Chem 9:644–653CrossRefPubMedGoogle Scholar
  69. 69.
    Toy PH, Newcomb M, Coon MJ, Vaz ADN (1998) Two distinct electrophilic oxidants effect hydroxylation in cytochrome P-450-catalyzed reactions. J Am Chem Soc 120:9718–9719CrossRefGoogle Scholar
  70. 70.
    Newcomb M, Le Tadic-Biadatti MH, Chestney DL, Roberts ES, Hollenberg PF (1995) A nonsynchronous concerted mechanism for cytochrome P-450 catalyzed hydroxylation. J Am Chem Soc 117:12085–12091CrossRefGoogle Scholar
  71. 71.
    Newcomb M, Aebisher D, Shen R, Chandrasena REP, Hollenberg PF, Coon MJ (2003) Kinetic isotope effects implicate two electrophilic oxidants in cytochrome P450-catalyzed hydroxylations. J Am Chem Soc 125:6064–6065CrossRefPubMedGoogle Scholar
  72. 72.
    Chandrasena REP, Vatsis KP, Coon MJ, Hollenberg PF, Newcomb M (2003) Hydroxylation by the hydroperoxy-iron species in cytochrome P450 enzymes. J Am Chem Soc 126:115–126CrossRefGoogle Scholar
  73. 73.
    Toy PH, Dhanabalasingam B, Newcomb M, Hanna IH, Hollenberg PF (1997) A substituted hypersensitive radical probe for enzyme-catalyzed hydroxylations: synthesis of racemic and enantiomerically enriched forms and application in a cytochrome P450-catalyzed oxidation. J Org Chem 62:9114–9122CrossRefGoogle Scholar
  74. 74.
    Ortiz de Montellano PR, Wilks A (2000) Advances in inorganic chemistry. In: Sykes G, Mauk AG (eds) Iron porphyrins, vol 51. Academic, San Diego, pp 359–407Google Scholar
  75. 75.
    Tenhunen R, Marver H, Pinstone NR, Trager WF, Cooper DY, Schmid R (1972) Enzymic degradation of heme. Oxygenative cleavage requiring cytochrome P-450. Biochemistry 11:1716–1720CrossRefPubMedGoogle Scholar
  76. 76.
    Wilks A, Ortiz de Montellano PR (1993) Rat liver heme oxygenase. High level expression of a truncated soluble form and nature of the meso-hydroxylating species. J Biol Chem 268:22357–22362PubMedGoogle Scholar
  77. 77.
    Wilks A, Torpey J, Ortiz de Montellano PR (1994) Heme oxygenase (HO-1). Evidence for electrophilic oxygen addition to the porphyrin ring in the formation of α-meso-hydroxyheme. J Biol Chem 269:29553–29556PubMedGoogle Scholar
  78. 78.
    Sundaramoorthy M, Terner J, Poulos TL (1995) The crystal structure of chloroperoxidase: a heme peroxidase–cytochrome P450 functional hybrid. Structure 3:1367–1378CrossRefPubMedGoogle Scholar
  79. 79.
    Groves JT, Wang CCY (2000) Nitric oxide synthase: models and mechanisms. Curr Opin Chem Biol 4:687–695CrossRefPubMedGoogle Scholar
  80. 80.
    Makris TM, Schlichting I, Sligar SG (2005) Activation of molecular oxygen by cytochrome P450. In: Ortiz de Montellano PR (ed) Cytochrome P450: structure, mechanism, and biochemistry, 2nd edn. Plenum Press, New York, pp 149–182CrossRefGoogle Scholar
  81. 81.
    Groves JT (2003) The bioinorganic chemistry of iron in oxygenases and supramolecular assemblies. Proc Natl Acad Sci U S A 100:3569–3574CrossRefPubMedCentralPubMedGoogle Scholar
  82. 82.
    Groves JT, Haushalter RC, Nakamura M, Nemo TE, Evans BJ (1981) High-valent iron-porphyrin complexes related to peroxidase and cytochrome P-450. J Am Chem Soc 103:2884–2886CrossRefGoogle Scholar
  83. 83.
    Groves JT, McClusky GA (1976) Aliphatic hydroxylation via oxygen rebound. Oxygen transfer catalyzed by iron. J Am Chem Soc 98:859–861CrossRefGoogle Scholar
  84. 84.
    Groves JT, Watanabe Y (1988) Reactive iron porphyrin derivatives related to the catalytic cycles of cytochrome P-450 and peroxidase. Studies of the mechanism of oxygen activation. J Am Chem Soc 110:8443–8452CrossRefGoogle Scholar
  85. 85.
    Shapiro S, Piper JU, Caspi E (1982) Steric course of hydroxylation at primary carbon atoms. Biosynthesis of 1-octanol from (1R)- and (1S)-[1-3H, 2H, 1H; 1-14C]octane by rat liver microsomes. J Am Chem Soc 104:2301–2305CrossRefGoogle Scholar
  86. 86.
    Shaik S, Filatov M, Schröder D, Schwarz H (1998) Electronic structure makes a difference: cytochrome P-450 mediated hydroxylations of hydrocarbons as a two-state reactivity paradigm. Chem Eur J 4:193–199CrossRefGoogle Scholar
  87. 87.
    Harris N, Cohen S, Filatov M, Ogliaro F, Shaik S (2000) Two-state reactivity in the rebound step of alkane hydroxylation by cytochrome P-450: origins of free radicals with finite lifetimes. Angew Chem Int Ed 39:2003–2007CrossRefGoogle Scholar
  88. 88.
    Ogliaro F, de Visser SP, Groves JT, Shaik S (2001) Chameleon states: high-valent metal-oxo species of cytochrome P450 and its ruthenium analogue. Angew Chem Int Ed 40:2874–2878CrossRefGoogle Scholar
  89. 89.
    Ogliaro F, Harris N, Cohen S, Filatov M, de Visser SP, Shaik S (2000) A model “rebound” mechanism of hydroxylation by cytochrome P450: stepwise and effectively concerted pathways, and their reactivity patterns. J Am Chem Soc 122:8977–8989CrossRefGoogle Scholar
  90. 90.
    Krest CM, Onderko EL, Yosca TH, Calixto JC, Karp RF, Livada J, Rittle J, Green MT (2013) Reactive intermediates in cytochrome P450 catalysis. J Biol Chem 288:17074–17081CrossRefPubMedCentralPubMedGoogle Scholar
  91. 91.
    Cryle MJ, De Voss JJ (2006) Is the ferric hydroperoxy species responsible for sulfur oxidation in cytochrome P450s? Angew Chem Int Ed 45:8221–8223CrossRefGoogle Scholar
  92. 92.
    Truan G, Komandla MR, Falck JR, Peterson JA (1999) P450BM-3: absolute configuration of the primary metabolites of palmitic acid. Arch Biochem Biophys 366:192–198CrossRefPubMedGoogle Scholar
  93. 93.
    Capdevila JH, Wei S, Helvig C, Falck JR, Belosludtsev Y, Truan G, Graham-Lorence SE, Peterson JA (1996) The highly stereoselective oxidation of polyunsaturated fatty acids by cytochrome P450BM-3. J Biol Chem 271:22663–22671CrossRefPubMedGoogle Scholar
  94. 94.
    Wang B, Li C, Cho KB, Nam W, Shaik S (2013) The FeIII(H2O2) complex as a highly efficient oxidant in sulfoxidation reactions: revival of an underrated oxidant in cytochrome P450. J Chem Theory Comput 9:2519–2525CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Department of Chemistry and BiochemistryUniversity of South CarolinaColumbiaUSA

Personalised recommendations