Annexin 2-caveolin 1 complex is a target of ezetimibe and regulates intestinal cholesterol transport (Retracted article. See vol. 110, pg. 797, 2013)
Annexin 2-caveolin 1 complex is a target of ezetimibe and regulates intestinal cholesterol transport (Retracted article. See vol. 110, pg. 797, 2013)
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DOI：
10.1073/pnas.0400441101
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发表时间：
2004-03-09
影响因子：
11.1
通讯作者：
Farber, SA
中科院分区：
文献类型：
--
作者：
Smart, EJ;De Rose, RA;Farber, SA
肠道是人体摄取膳食胆固醇的主要场所，调节肠道中胆固醇的吸收具有深远的临床意义。我们已经进行了反向遗传的方法，通过破坏假定的胆固醇加工基因在斑马鱼幼虫中使用吗啉代（MO）反义寡核苷酸。通过使用靶向MO注射和免疫沉淀（IP）实验结合质谱，我们确定了膜联蛋白（ANX）2与小窝蛋白（CAV）1在斑马鱼和小鼠肠道中的复合物。该复合物是热稳定的，不受SDS或还原条件的影响。MO靶向anx2b或cav1，这两者都强烈表达在幼虫和成年斑马鱼肠上皮细胞，防止蛋白质杂合物的形成。此外，anx2b MO注射防止荧光胆固醇报告分子的加工，并导致甾醇质量降低。依折麦布对小鼠的药理学处理仅破坏了高胆固醇血症动物中的杂合物。这些数据表明ANX2和CAV 1是肠固醇转运复合物的组分。
Modulation of cholesterol absorption in the intestine, the primary site of dietary cholesterol uptake in humans, can have profound clinical implications. We have undertaken a reverse genetic approach by disrupting putative cholesterol processing genes in zebrafish larvae by using morpholino (MO) antisense oligonucleoticles. By using targeted MO injections and immunoprecipitation (IP) experiments coupled with mass spectrometry, we determined that annexin (ANX)2 complexes with caveolin (CAV)1 in the zebrafish and mouse intestine. The complex is heat stable and unaffected by SDS or reducing conditions. MO targeting of anx2b or cav1, which are both strongly expressed in the larval and adult zebrafish intestinal epithelium, prevents formation of the protein heterocomplex. Furthermore, anx2b MO injection prevents processing of a fluorescent cholesterol reporter and results in reduced sterol mass. Pharmacological treatment of mice with ezetimibe disrupts the heterocomplex in only hypercholesterolemic animals. These data suggest that ANX2 and CAV1 are components of an intestinal sterol transport complex.