Analysis of Cyclooxygenase-Substrate Interactions Using Stereospecificallylabeled Arachidonic Acids

  • Claus Schneider
  • William E. Boeglin
  • Alan R. Brash
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 507)


The two mammalian cyclooxygenase isoforms, COX-1 and COX-2, catalyze an identical reaction in the formation of prostaglandin H2 from arachidonic acid (Smith et al., 1996). The initial abstraction of the pro-S hydrogen at carbon C-13 is followed by oxygenation in the 11R position, formation of the endoperoxide and the five-membered prostaglandin ring, and, finally, a second oxygenation at C-15 to yield prostaglandin H2 (Hamberg and Samuelsson, 1967a).

As with all cyclooxygenases and lipoxygenases investigated so far, this mechanism follows the so-called antarafacial rule of hydrogen abstraction and oxygen insertion: both events occur on opposite sites of the planar cis,cis-pentadiene system of arachidonic acid (Hamberg and Samuelsson, 1967a; 1967b). COX-1 and COX-2, however, are different in their response to treatment with the non-steroidal anti-inflammatory drug, aspirin. In both enzymes, aspirin acetylates a critical serine residue in the active site channel. This leads to a complete block of oxygenase activity in COX-1, but to a new oxygenase specificity in COX-2, namely, formation of 15R-hydroxyeicosatetraenoic acid (15R-HETE) as the sole enzymatic product (Holtzman et al., 1992; Meade et al., 1993; Lecomte et al., 1994).


Arachidonic Acid Hydrogen Abstraction Binding Conformation Prostaglandin Endoperoxide Octadecatrienoic Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Hamberg, M. and Samuelsson, B., 1967a, Oxygenation of unsaturated fatty acids by the vesicular gland of sheepJ. Biol. Chem.242:5344.PubMedGoogle Scholar
  2. 2.
    Hamberg, M. and Samuelsson, B., 1967b, On the specificity of the oxygenation of unsaturated fatty acids catalyzed by soybean lipoxidaseJ. Biol. Chem.242:5329.PubMedGoogle Scholar
  3. 3.
    Holtzman, M.J., Turk, J., and Shornick, L.P., 1992, Identification of a pharmacologically distinct prostaglandin H synthase in cultured epithelial cellsJ. Biol. Chem.267:21438.PubMedGoogle Scholar
  4. 4.
    Kurumbail, R.G., Stevens, A.M., Gierse, J.K., McDonald, J.J., Stegeman, R.A., Pak, J.Y., Gildehaus, J.M., Miyashiro, J.M., Penning, T.D., Seibert, K., Isakson, P.C., and Stallings, W.C., 1996, Structural basis for selective inhibition of cyclooxygenase-2 by anti-inflammatory agentsNature384:644.PubMedCrossRefGoogle Scholar
  5. 5.
    Laneuville, O., Breuer, D.K., Xu, N., Huang, Z.H., Gage, D.A., Watson, J.T., Lagarde, M., DeWitt, D.L., and Smith, W.L., 1995, Fatty acid substrate specificities of human prostaglandin-endoperoxide H synthase-1 and -2. Formation of t2-hydroxy-(9Z, I 3E/Z, 15Z)-octadecatrienoic acids from alphalinolenic acidJ. Biol. Chem.270:19330.PubMedCrossRefGoogle Scholar
  6. 6.
    Lecomte, M., Laneuville, O., Ji, C., DeWitt, D.L., and Smith, W.L., 1994, Acetylation of human prostaglandin endoperoxide synthase-2 (cyclooxygenase-2) by aspirinJ. Biol. Chem.269:13207.PubMedGoogle Scholar
  7. 7.
    Luong, C., Miller, A., Barnett, J., Chow, J., Ramesha, C., and Browner, M.F., 1996, Flexibility of the NSAID binding site in the structure of human cyclooxygenase-2Nat. Struct. Biol.3:927.PubMedCrossRefGoogle Scholar
  8. 8.
    Meade, E.A., Smith, W.L., and DeWitt, D.L., 1993, Differential inhibition of prostaglandin endoperoxide synthase (cyclooxygenase) isoenzymes by aspirin and other non-steroidal anti-inflammatory drugsJ. Biol. Chem.268: 6610.PubMedGoogle Scholar
  9. 9.
    Rowlinson, S.W., Crews, B.C., Lanzo, C.A., and Marnett, L.J., 1999, The binding of arachidonic acid in the cyclo-oxygenase active site of mouse prostaglandin endoperoxide synthase-2 (COX-2). A putative L-shaped binding conformation utilizing the top channel regionJ. Biol. Chem.274:23305.PubMedCrossRefGoogle Scholar
  10. 10.
    Rowlinson, S.W., Crews, B.C., Goodwin, D.C., Schneider, C., Gierse, J.K., and Marnett, L.J., 2000, Spatial require-ments for 15-(R)-Hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic acid synthesis within the cyclooxygenase active site of murine COX-2J. Biol. Chem.275:6586.PubMedCrossRefGoogle Scholar
  11. 11.
    Schneider, C. and Brash, A.R., 2000, Stereospecificity of hydrogen abstraction in the conversion of arachidonic acid to 15R-HETE by aspirin-treated cyclooxygenase-2. Implications for the alignment of substrate in the active siteJ. Biol. Chem.275:4743.PubMedCrossRefGoogle Scholar
  12. 12.
    Smith, W.L., Garavito, R.M., and DeWitt, D.L., 1996, Prostaglandin endoperoxide H synthases (cyclooxygenases)-1 and —2J. Biol. Chem.271:33157.PubMedCrossRefGoogle Scholar
  13. 13.
    Tang, M.S., Copeland, R.A., and Penning, T.M., 1997, Detection of an Fe“-protoporphyrin-IX intermediate during aspirin-treated prostaglandin H2 synthase II catalysis of arachidonic acid to 15HETEBiochemistry36: 7527.PubMedCrossRefGoogle Scholar
  14. 14.
    Xiao G., Tsai A.L., Palmer G., Boyar W.C., Marshall P.J., Kulmacz R.J., 1997, Analysis of hydroperoxide-induced tyrosyl radicals and lipoxygenase activity in aspirin-treated human prostaglandin H synthase-2Biochemistry36:1836.PubMedCrossRefGoogle Scholar
  15. 15.
    Yu, M., Ives, D., and Ramesha, C.S., 1997, Synthesis of prostaglandin E2 ethanolamide from anandamide by cyclo-oxygenase-2J. Biol. Chem.272:21181.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Claus Schneider
    • 1
  • William E. Boeglin
    • 1
  • Alan R. Brash
    • 1
  1. 1.Department of PharmacologyVanderbilt University School of MedicineNashvilleUSA

Personalised recommendations