Platelet-Type Arachidonate 12-Lipoxygenase

  • Michihiro Nakamura
  • Natsuo Ueda
  • Shozo Yamamoto
  • Kazunori Ishimura
  • Kenjiro Tomo
  • Minoru Okuma
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 416)


In human platelets arachidonic acid is released from membrane phospholipids by the catalysis of phosopholipase A2, and then oxygenated either by cyclooxygenase or by 12-lipoxygenase. It is well known that the cyclooxygenase pathway leads to the synthesis of pro-aggregatory and vasoconstrictive thromboxane A2. 12-Lipoxygenase introduces one oxygen molecule regiospecifically and stereoselectively into the carbon 12 of arachidonic acid. The primary product is 12S-hydroperoxy-5, 8, 14-cis-l0-trans-eicosatetraenoic acid (12-HPETE), which is reduced to 12S-hydroxy acid (12-HETE) in a whole cell preparation. A number of papers have reported biological activities of either 12-HPETE or 12-HETE1. However, most of them could not be generalized in terms of the cell type and animal species, and the general biological significance of the enzyme has not been well understood1,2. Recently two groups of bioactive arachidonate metabolites were identified as the products of the 12-lipoxygenase pathway. Namely, transcellular synthesis of lipoxins via leukotriene A4 requires the participation of platelet 12-lipoxygenase3, and 12-HPETE is enzymatically transformed to hepoxilins4. Our group has demonstrated the occurrence of two distinct isoforms of l2-lipoxygenase in mammalian tissues1,5. The leukocyte-type 12-lipoxygenase has a broad substrate specificity reacting with linoleic and linolenic acids with 18 carbon atoms as well as arachidonic acid with 20 carbon atoms. In contrast, the platelet-type enzyme is much less active with the C18 fatty acids. The leukocyte-type enzyme is also found in other tissues in addition to leukocytes1. The platelet-type enzyme was found earlier in platelets of various animal species, and found recently in epidermis6–8. For a better understanding of the physiological role of the platelet-type 12-lipoxygenase we planned immunohistochemical studies of the enzyme. However, purification of this enzyme from platelets was difficult, and no polyclonal antibody with a high affinity for the enzyme had not been available until recently. Therefore, we attempted to purify a recombinant enzyme as antigen.


Differential Centrifugation Subcellular Organelle Mouse Bone Marrow Cell Human Blood Platelet Electron Microscopic Immunocytochemistry 
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Copyright information

© Springer Science+Business Media New York 1996

Authors and Affiliations

  • Michihiro Nakamura
    • 1
  • Natsuo Ueda
    • 1
  • Shozo Yamamoto
    • 1
  • Kazunori Ishimura
    • 2
  • Kenjiro Tomo
    • 3
  • Minoru Okuma
    • 3
  1. 1.Department of Biochemistry School of MedicineTokushima UniversityTokushima 770Japan
  2. 2.Department of Anatomy School of MedicineTokushima UniversityTokushima 770Japan
  3. 3.The First Division, Department of Internal Medicine Faculty of MedicineKyoto UniversityKyoto 606-01Japan

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