Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells.

Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Research Abstract Details 

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Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Abstract Text:

Karl-Rudolf Erlemann,Chantal Cossette,Gail E Grant,Gue-Jae Lee,Pranav Patel,Joshua Rokach,William S Powell,

The 5-lipoxygenase product 5-oxo-ETE (5-oxo-eicosatetraenoic acid) is a highly potent granulocyte chemoattractant that is synthesized from 5-HETE (5-hydroxyeicosatetraenoic acid) by 5-HEDH (5-hydroxyeicosanoid dehydrogenase). In the present study, we found that 5-HEDH activity is induced in U937 monocytic cells by differentiation towards macrophages with PMA and in HL-60 myeloblastic cells by 1,25-dihydroxy-vitamin D3. We used PMA-differentiated U937 cells to investigate further the regulation of 5-HEDH. This enzyme exhibits approx. 10000-fold selectivity for NADP+ over NAD+ as a cofactor for the oxidation of 5-HETE, which is maximal at pH 10.2. In contrast, the reverse reaction (5-oxo-ETE-->5-HETE) is NADPH-dependent and is maximal at pH 6. Although the K(m) for the forward reaction (670 nM) is about twice that for the reverse reaction at neutral pH, the V(max) is approx 8-fold higher. The oxidation of 5-HETE to 5-oxo-ETE is supported by very low concentrations of NADP(+) (K(m) 139 nM), inhibited by NADPH (K(i) 224 nM) and is consistent with a ping-pong mechanism. The amount of 5-oxo-ETE synthesized by 5-HEDH depends on the ratio of NADP+ to NADPH. Exposure of U937 cells to oxidative stress (t-butyl hydroperoxide) increased the ratio of NADP+ to NADPH from approx. 0.08 in resting cells to approx. 3, and this was accompanied by a dramatic increase in 5-HETE oxidation to 5-oxo-ETE. We conclude that differentiation of monocytic cells towards macrophages results in enhanced 5-oxo-ETE synthesis and that the ability of cells to synthesize 5-oxo-ETE is tightly regulated by the ratio of intracellular NADP+ to NADPH.

Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Publishing Authors By Initials

KR Erlemann,C Cossette,GE Grant,GJ Lee,P Patel,J Rokach,WS Powell,

For similar cells: cells, cultured: cell line: cell line, tumor: u937 cells research abstracts see: cells: cells, cultured: cell line: cell line, tumor: u937 cells research

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Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: The Biochemical journal

VOLUME: 403

Page Numbers: 157-65

Journal Abbreviation: Biochem. J.

ISSN: 1470-8728

DAY: 1

MONTH: Apr

YEAR: 2007

Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 2984726

Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Keywords Mesh Terms:

KEYWORDS: U937 Cells

MESH TERMS: pharmacology

Chemical & Substance for Abstract: Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells. Information

Substance Name: 5-hydroxyeicosanoid dehydrogenase

Registry Number: EC 1.1.1.-

Grant and Affiliation Information for Regulation of 5-hydroxyeicosanoid dehydrogenase activity in monocytic cells.

AFFILIATION: Meakins-Christie Laboratories, Department of Medicine, McGill University, 3626 St. Urbain Street, Montreal, Quebec H2X 2P2, Canada.

Country: England

AGENCY: United States NHLBI

GRANT: HL81873

ACRONYM: HL

MEDLINETA: Biochem J

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