Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells
Okadaic acid was found to induce concentration- and time-dependent lipolysis in rat fat cells in the absence of lipolytic hormones, but it did not significantly increase the total hormone-sensitive lipase (HSL) activity in these fat cells, the activity of HSL extracted from fat layer and that of HSL...
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Elsevier
2000-02-01
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Series: | Journal of Lipid Research |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520320538 |
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author | Chie Morimoto Atsuko Kiyama Kenji Kameda Hiroyuki Ninomiya Takahiro Tsujita Hiromichi Okuda |
author_facet | Chie Morimoto Atsuko Kiyama Kenji Kameda Hiroyuki Ninomiya Takahiro Tsujita Hiromichi Okuda |
author_sort | Chie Morimoto |
collection | DOAJ |
description | Okadaic acid was found to induce concentration- and time-dependent lipolysis in rat fat cells in the absence of lipolytic hormones, but it did not significantly increase the total hormone-sensitive lipase (HSL) activity in these fat cells, the activity of HSL extracted from fat layer and that of HSL in the supernatant of homogenized fat cells. Western blotting of fat cell homogenate fractions with an antiserum raised against synthetic peptide derived from rat HSL showed that HSL protein shifted from the supernatant to the fat layer in response to okadaic acid, which increased the HSL protein content on the fat layer and concomitantly reduced that of the supernatant, concentration- and time-dependently. Sonication of the fat cells abolished their responsiveness to okadaic acid. The lipolytic action of okadaic acid was examined and its site was identified using a cell-free system comprising lipid droplets isolated from rat fat cells and HSL. Okadaic acid induced lipolysis in this cell-free system and sonication of the lipid droplets caused disappearance of lipolytic action of okadaic acid. Okadaic acid failed to stimulate lipolysis in a cell-free system comprising HSL and artificial lipid droplets (trioleoylglycerol emulsified with gum arabic) instead of lipid droplets isolated from rat fat cells. These results suggest that okadaic acid does not increase the catalytic activity of HSL but induces translocation of HSL to the lipid droplets isolated from rat fat cells. The site of the lipolytic action of okadaic acid in relation to the interaction between HSL and lipid droplet is discussed. —Morimoto, C., A. Kiyama, K. Kameda, H. Ninomiya, T. Tsujita, and H. Okuda. Mechanism of the stimulatory action of okadic acid on lipolysis in rat fat cells. |
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issn | 0022-2275 |
language | English |
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spelling | doaj.art-d88891ff80d24ab8973a96ace3ad72ed2022-12-21T21:30:19ZengElsevierJournal of Lipid Research0022-22752000-02-01412199204Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cellsChie Morimoto0Atsuko Kiyama1Kenji Kameda2Hiroyuki Ninomiya3Takahiro Tsujita4Hiromichi Okuda52nd Department of Medical Biochemistry, School of Medicine, Ehime University, Shigenobu-cho, Onsen-gun, Ehime 791-0295; Department of Chemistry, Faculty of Science, Ehime University, Matsuyama, Ehime 790-8577Department of Human and Natural Environment Sciences, Faculty of Integrated Arts and Sciences, Tokushima University, Minamijousanjima-cho, Tokushima 770-8502, JapanCentral Research Laboratory, School of Medicine, Ehime University, Shigenobu-cho, Onsen-gun, Ehime 791-02952nd Department of Medical Biochemistry, School of Medicine, Ehime University, Shigenobu-cho, Onsen-gun, Ehime 791-0295Central Research Laboratory, School of Medicine, Ehime University, Shigenobu-cho, Onsen-gun, Ehime 791-0295To whom correspondence should be addressed.; 2nd Department of Medical Biochemistry, School of Medicine, Ehime University, Shigenobu-cho, Onsen-gun, Ehime 791-0295Okadaic acid was found to induce concentration- and time-dependent lipolysis in rat fat cells in the absence of lipolytic hormones, but it did not significantly increase the total hormone-sensitive lipase (HSL) activity in these fat cells, the activity of HSL extracted from fat layer and that of HSL in the supernatant of homogenized fat cells. Western blotting of fat cell homogenate fractions with an antiserum raised against synthetic peptide derived from rat HSL showed that HSL protein shifted from the supernatant to the fat layer in response to okadaic acid, which increased the HSL protein content on the fat layer and concomitantly reduced that of the supernatant, concentration- and time-dependently. Sonication of the fat cells abolished their responsiveness to okadaic acid. The lipolytic action of okadaic acid was examined and its site was identified using a cell-free system comprising lipid droplets isolated from rat fat cells and HSL. Okadaic acid induced lipolysis in this cell-free system and sonication of the lipid droplets caused disappearance of lipolytic action of okadaic acid. Okadaic acid failed to stimulate lipolysis in a cell-free system comprising HSL and artificial lipid droplets (trioleoylglycerol emulsified with gum arabic) instead of lipid droplets isolated from rat fat cells. These results suggest that okadaic acid does not increase the catalytic activity of HSL but induces translocation of HSL to the lipid droplets isolated from rat fat cells. The site of the lipolytic action of okadaic acid in relation to the interaction between HSL and lipid droplet is discussed. —Morimoto, C., A. Kiyama, K. Kameda, H. Ninomiya, T. Tsujita, and H. Okuda. Mechanism of the stimulatory action of okadic acid on lipolysis in rat fat cells.http://www.sciencedirect.com/science/article/pii/S0022227520320538okadaic acidlipolysisHSL activityfat cellendogenous lipid droplet |
spellingShingle | Chie Morimoto Atsuko Kiyama Kenji Kameda Hiroyuki Ninomiya Takahiro Tsujita Hiromichi Okuda Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells Journal of Lipid Research okadaic acid lipolysis HSL activity fat cell endogenous lipid droplet |
title | Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells |
title_full | Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells |
title_fullStr | Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells |
title_full_unstemmed | Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells |
title_short | Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells |
title_sort | mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells |
topic | okadaic acid lipolysis HSL activity fat cell endogenous lipid droplet |
url | http://www.sciencedirect.com/science/article/pii/S0022227520320538 |
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