Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells

Abstract Beauveriolide III (BeauIII) inhibited sterol O-acyltransferases 1 and 2 (SOAT1 and SOAT2), which are endoplasmic reticulum (ER) membrane proteins, in an enzyme-based assay, and selectively inhibited SOAT1 in a cell-based assay using SOAT1-/SOAT2-CHO cells. This discrepancy in SOAT inhibitio...

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Autores principales:	Taichi Ohshiro, Keisuke Kobayashi, Mio Ohba, Daisuke Matsuda, Lawrence L. Rudel, Takashi Takahashi, Takayuki Doi, Hiroshi Tomoda
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ac9b0bfcc18c4273ac51f31d3d43296a
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spelling	oai:doaj.org-article:ac9b0bfcc18c4273ac51f31d3d43296a2021-12-02T11:53:10ZSelective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells10.1038/s41598-017-04177-82045-2322https://doaj.org/article/ac9b0bfcc18c4273ac51f31d3d43296a2017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04177-8https://doaj.org/toc/2045-2322Abstract Beauveriolide III (BeauIII) inhibited sterol O-acyltransferases 1 and 2 (SOAT1 and SOAT2), which are endoplasmic reticulum (ER) membrane proteins, in an enzyme-based assay, and selectively inhibited SOAT1 in a cell-based assay using SOAT1-/SOAT2-CHO cells. This discrepancy in SOAT inhibition by BeauIII was investigated. In the enzyme-based assay, BeauIII inhibited SOAT1 and SOAT2 to a similar extent using microsomes prepared from cells disrupted under the strongest sonication condition. In semi-intact SOAT1-/SOAT2-CHO cells prepared by a treatment with digitonin (plasma membrane permeabilized), BeauIII selectively inhibited SOAT1 (IC50; 5.0 µM (SOAT1) vs >90 µM (SOAT2)), while in those treated with saponin (plasma membrane and ER membrane permeabilized), BeauIII inhibited SOAT1 (IC50, 1.8 µM) and SOAT2 (5.9 µM). SOAT1-selective inhibition by BeauIII was reproduced in intact ER fractions prepared from SOAT1/SOAT2-CHO cells. A Western blotting analysis revealed that biotin-labeled beauveriolide bound to the SOAT1 protein prepared from SOAT1-CHO cells. We concluded that BeauIII binds to a putative active site responsible for SOAT1 that is located on the cytosolic side of the ER, while BeauIII is not accessible to the corresponding active site for SOAT2 located on the luminal side.Taichi OhshiroKeisuke KobayashiMio OhbaDaisuke MatsudaLawrence L. RudelTakashi TakahashiTakayuki DoiHiroshi TomodaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Taichi Ohshiro Keisuke Kobayashi Mio Ohba Daisuke Matsuda Lawrence L. Rudel Takashi Takahashi Takayuki Doi Hiroshi Tomoda Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells
description	Abstract Beauveriolide III (BeauIII) inhibited sterol O-acyltransferases 1 and 2 (SOAT1 and SOAT2), which are endoplasmic reticulum (ER) membrane proteins, in an enzyme-based assay, and selectively inhibited SOAT1 in a cell-based assay using SOAT1-/SOAT2-CHO cells. This discrepancy in SOAT inhibition by BeauIII was investigated. In the enzyme-based assay, BeauIII inhibited SOAT1 and SOAT2 to a similar extent using microsomes prepared from cells disrupted under the strongest sonication condition. In semi-intact SOAT1-/SOAT2-CHO cells prepared by a treatment with digitonin (plasma membrane permeabilized), BeauIII selectively inhibited SOAT1 (IC50; 5.0 µM (SOAT1) vs >90 µM (SOAT2)), while in those treated with saponin (plasma membrane and ER membrane permeabilized), BeauIII inhibited SOAT1 (IC50, 1.8 µM) and SOAT2 (5.9 µM). SOAT1-selective inhibition by BeauIII was reproduced in intact ER fractions prepared from SOAT1/SOAT2-CHO cells. A Western blotting analysis revealed that biotin-labeled beauveriolide bound to the SOAT1 protein prepared from SOAT1-CHO cells. We concluded that BeauIII binds to a putative active site responsible for SOAT1 that is located on the cytosolic side of the ER, while BeauIII is not accessible to the corresponding active site for SOAT2 located on the luminal side.
format	article
author	Taichi Ohshiro Keisuke Kobayashi Mio Ohba Daisuke Matsuda Lawrence L. Rudel Takashi Takahashi Takayuki Doi Hiroshi Tomoda
author_facet	Taichi Ohshiro Keisuke Kobayashi Mio Ohba Daisuke Matsuda Lawrence L. Rudel Takashi Takahashi Takayuki Doi Hiroshi Tomoda
author_sort	Taichi Ohshiro
title	Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells
title_short	Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells
title_full	Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells
title_fullStr	Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells
title_full_unstemmed	Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells
title_sort	selective inhibition of sterolo-acyltransferase 1 isozyme by beauveriolide iii in intact cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/ac9b0bfcc18c4273ac51f31d3d43296a
work_keys_str_mv	AT taichiohshiro selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT keisukekobayashi selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT mioohba selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT daisukematsuda selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT lawrencelrudel selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT takashitakahashi selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT takayukidoi selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells AT hiroshitomoda selectiveinhibitionofsteroloacyltransferase1isozymebybeauveriolideiiiinintactcells
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Selective inhibition of sterolO-acyltransferase 1 isozyme by beauveriolide III in intact cells

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