Leukotriene b ω-hydroxylase in rat liver microsomes: Identification as a1 cytochrome P-450 that catalyzes prostaglandin a ω-hydroxylation, and participation of cytochrome b5

Hideki Sumimoto, Emi Kusunose, Yoichi Gotoh, Masamichi Kusunose, Shigeki Minakami

Research output: Contribution to journalArticle

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Abstract

The ω-hydroxylation of leukotriene B4 (LTB4 by rat liver microsomes requires NADPH and molecular oxygen, suggesting that the hydroxylation is catalyzed by a cytochrome P-450 (P-450)-linked monooxygenase system. The reaction is inhibited by CO, and the inhibition is reversed by irradiation of light at 450 nm in a light-intensity-dependent manner. The extent of the reversal is strongly dependent on the wavelength of the light used, the 450-nm light is most efficient. The finding provides direct evidence for the identification of the LTB ω-hydroxylase as a P-450. The P-450 seems to be also responsible for prostaglandin A, (PGA aj-hydroxylation, but not for laurie aicd ω-hydroxylation. The LTB. ω-hydroxylation is competitively inhibited by PGAI, but not affected by lauric acid. The K, value for PGAI of 38 M agrees with the X value for PGA ω-hydroxylation of 40 μM. LTB inhibits the PGAI ω-hydroxylation by rat liver microsomes in a competitive manner with the K, of 43 pM, which is consistent with the K for the LTB ω-hydroxylation of 42 μM. An antiserum raised against rabbit pulmonary PG ω-hydroxylase (P-450p-2 inhibits slightly the ω-hydroxylations of LTB and PGAI, while it has stronger inhibitory effect on lauric acid ω-hydroxylation. In addition to NADPH-cytochrome P-450 reductase, cytochrome b5 appears to participate in the LTB ω-hydroxylating system, since the reaction is inhibited by an antibody raised against the cytochrome b as well as one raised against the reductase.

Original languageEnglish
Pages (from-to)215-221
Number of pages7
JournalJournal of biochemistry
Volume108
Issue number2
DOIs
Publication statusPublished - Jan 1 1990

Fingerprint

Cytochromes b5
Hydroxylation
Leukotrienes
Liver Microsomes
Mixed Function Oxygenases
Liver
Cytochrome P-450 Enzyme System
Prostaglandins
Rats
lauric acid
Prostaglandins A
Light
Leukotriene B4
NADPH-Ferrihemoprotein Reductase
Cytochromes b
Molecular oxygen
Carbon Monoxide
NADP
Immune Sera
Oxidoreductases

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Leukotriene b ω-hydroxylase in rat liver microsomes : Identification as a1 cytochrome P-450 that catalyzes prostaglandin a ω-hydroxylation, and participation of cytochrome b5. / Sumimoto, Hideki; Kusunose, Emi; Gotoh, Yoichi; Kusunose, Masamichi; Minakami, Shigeki.

In: Journal of biochemistry, Vol. 108, No. 2, 01.01.1990, p. 215-221.

Research output: Contribution to journalArticle

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abstract = "The ω-hydroxylation of leukotriene B4 (LTB4 by rat liver microsomes requires NADPH and molecular oxygen, suggesting that the hydroxylation is catalyzed by a cytochrome P-450 (P-450)-linked monooxygenase system. The reaction is inhibited by CO, and the inhibition is reversed by irradiation of light at 450 nm in a light-intensity-dependent manner. The extent of the reversal is strongly dependent on the wavelength of the light used, the 450-nm light is most efficient. The finding provides direct evidence for the identification of the LTB ω-hydroxylase as a P-450. The P-450 seems to be also responsible for prostaglandin A, (PGA aj-hydroxylation, but not for laurie aicd ω-hydroxylation. The LTB. ω-hydroxylation is competitively inhibited by PGAI, but not affected by lauric acid. The K, value for PGAI of 38 M agrees with the X value for PGA ω-hydroxylation of 40 μM. LTB inhibits the PGAI ω-hydroxylation by rat liver microsomes in a competitive manner with the K, of 43 pM, which is consistent with the K for the LTB ω-hydroxylation of 42 μM. An antiserum raised against rabbit pulmonary PG ω-hydroxylase (P-450p-2 inhibits slightly the ω-hydroxylations of LTB and PGAI, while it has stronger inhibitory effect on lauric acid ω-hydroxylation. In addition to NADPH-cytochrome P-450 reductase, cytochrome b5 appears to participate in the LTB ω-hydroxylating system, since the reaction is inhibited by an antibody raised against the cytochrome b as well as one raised against the reductase.",
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N2 - The ω-hydroxylation of leukotriene B4 (LTB4 by rat liver microsomes requires NADPH and molecular oxygen, suggesting that the hydroxylation is catalyzed by a cytochrome P-450 (P-450)-linked monooxygenase system. The reaction is inhibited by CO, and the inhibition is reversed by irradiation of light at 450 nm in a light-intensity-dependent manner. The extent of the reversal is strongly dependent on the wavelength of the light used, the 450-nm light is most efficient. The finding provides direct evidence for the identification of the LTB ω-hydroxylase as a P-450. The P-450 seems to be also responsible for prostaglandin A, (PGA aj-hydroxylation, but not for laurie aicd ω-hydroxylation. The LTB. ω-hydroxylation is competitively inhibited by PGAI, but not affected by lauric acid. The K, value for PGAI of 38 M agrees with the X value for PGA ω-hydroxylation of 40 μM. LTB inhibits the PGAI ω-hydroxylation by rat liver microsomes in a competitive manner with the K, of 43 pM, which is consistent with the K for the LTB ω-hydroxylation of 42 μM. An antiserum raised against rabbit pulmonary PG ω-hydroxylase (P-450p-2 inhibits slightly the ω-hydroxylations of LTB and PGAI, while it has stronger inhibitory effect on lauric acid ω-hydroxylation. In addition to NADPH-cytochrome P-450 reductase, cytochrome b5 appears to participate in the LTB ω-hydroxylating system, since the reaction is inhibited by an antibody raised against the cytochrome b as well as one raised against the reductase.

AB - The ω-hydroxylation of leukotriene B4 (LTB4 by rat liver microsomes requires NADPH and molecular oxygen, suggesting that the hydroxylation is catalyzed by a cytochrome P-450 (P-450)-linked monooxygenase system. The reaction is inhibited by CO, and the inhibition is reversed by irradiation of light at 450 nm in a light-intensity-dependent manner. The extent of the reversal is strongly dependent on the wavelength of the light used, the 450-nm light is most efficient. The finding provides direct evidence for the identification of the LTB ω-hydroxylase as a P-450. The P-450 seems to be also responsible for prostaglandin A, (PGA aj-hydroxylation, but not for laurie aicd ω-hydroxylation. The LTB. ω-hydroxylation is competitively inhibited by PGAI, but not affected by lauric acid. The K, value for PGAI of 38 M agrees with the X value for PGA ω-hydroxylation of 40 μM. LTB inhibits the PGAI ω-hydroxylation by rat liver microsomes in a competitive manner with the K, of 43 pM, which is consistent with the K for the LTB ω-hydroxylation of 42 μM. An antiserum raised against rabbit pulmonary PG ω-hydroxylase (P-450p-2 inhibits slightly the ω-hydroxylations of LTB and PGAI, while it has stronger inhibitory effect on lauric acid ω-hydroxylation. In addition to NADPH-cytochrome P-450 reductase, cytochrome b5 appears to participate in the LTB ω-hydroxylating system, since the reaction is inhibited by an antibody raised against the cytochrome b as well as one raised against the reductase.

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