12(S)‐Hydroxyeicosatetraenoic acid induces cAMP production via increasing intracellular calcium concentration

FEBS Letters

Kumagai, Satoru¹  Hasegawa, Go¹  Yano, Masato¹  Saito, Yuji¹  Kobayashi, Yuichi²  Wang, Yong-Gang² 

[1] Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan;Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan

关键词: Hydroxyeicosatetraenoic acid;    Adenylyl cyclase;    Calcium;    cAMP;    12-HETE;    12(S)-hydroxy-5Z;    8Z;    10E;    14Z-eicosatetraenoic acid;    HETE;    hydroxyeicosatetraenoic acid;    AC;    adenylyl cyclase;    PMA;    phorbol 12-myristate 13-acetate;    IBMX;    3-isobutyl-1-methylxanthine;    W-13;    N-(4-aminobutyl)-5-chloro-2-naphthalenesulfonamide;    PGE1;    prostaglandin E1;    RT-PCR;    reverse transcriptase-polymerase chain reaction;   

DOI  :  10.1016/S0014-5793(03)01128-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have found that a 12-lipoxygenase metabolite of arachidonic acid, 12(S)-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-HETE), induces cAMP production in human normal fibroblast TIG-1 cells. This phenomenon was not observed in other cells tested including human embryonic kidney HEK293 cells. We have speculated that this specific response might be influenced by the kinds of isoform of adenylyl cyclase (AC) present in cells. We found that TIG-1 cells specifically expressed type VIII AC. As type VIII AC is known to be activated by an increase of calcium concentration, we determined the change of intracellular Ca²⁺ concentration after the addition of 12-HETE. It was elevated not only in TIG-1 cells, but also HEK293 cells, which did not respond to 12-HETE to produce cAMP. The addition of a calcium ionophore elevated the concentration of intracellular cAMP in TIG-1 cells, but it was without effect in HEK293 cells. To show that the expression of this particular isoform of AC is responsible for the positive response to 12-HETE, we transfected this AC isoform into HEK293 cells. The type VIII AC-transfected cells, in contrast to the mock-transfected ones, became very responsive to 12-HETE to produce cAMP. Taken all together the data would strongly suggest that 12-HETE specifically activates type VIII AC via increasing intracellular Ca²⁺ concentration.

【 授权许可】

Unknown