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

Taichi Ohshiro, Keisuke Kobayashi, Mio Ohba, Daisuke Matsuda, Lawrence L. Rudel, Takashi Takahashi, Takayuki Doi, Hiroshi Tomoda

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11 Citations (Scopus)

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.

Original languageEnglish
Article number4163
JournalScientific reports
Volume7
Issue number1
DOIs
Publication statusPublished - 2017 Dec 1

ASJC Scopus subject areas

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    Ohshiro, T., Kobayashi, K., Ohba, M., Matsuda, D., Rudel, L. L., Takahashi, T., Doi, T., & Tomoda, H. (2017). Selective inhibition of sterol O-acyltransferase 1 isozyme by beauveriolide III in intact cells. Scientific reports, 7(1), [4163]. https://doi.org/10.1038/s41598-017-04177-8