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首页> 外文期刊>Molecular biology of the cell >Essential role of Ca2+/calmodulin in early endosome antigen-1 localization
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Essential role of Ca2+/calmodulin in early endosome antigen-1 localization

机译：Ca2 + /钙调蛋白在早期内体抗原1定位中的重要作用

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摘要

Ca2+ is an essential requirement in membrane fusion, acting through binding proteins such as calmodulin (CaM). Ca2+/CaM is required for early endosome fusion in vitro, however, the molecular basis for this requirement is unknown. An additional requirement for endosome fusion is the protein Early Endosome Antigen 1 (EEA1), and its recruitment to the endosome depends on phosphatidylinositol 3-phosphate [PI(3)P] and the Rab5 GTPase. Herein, we demonstrate that inhibition of Ca2+/CaM, by using either chemical inhibitors or specific antibodies directed to CaM, results in a profound inhibition of EEA1 binding to endosomal membranes both in live cells and in vitro. The concentration of Ca2+/CaM inhibitors required for a full dissociation of EEA1 from endosomal membranes had no effect on the activity of phosphatidylinositol 3-kinases or on endogenous levels of PI(3)P. However, the interaction of EEA1 with liposomes containing PI(3)P was decreased by Ca2+/CaM inhibitors. Thus, Ca2+/CaM seems to be required for the stable interaction of EEA1 with endosomal PI(3)P, perhaps by directly or indirectly stabilizing the quaternary organization of the C-terminal FYVE domain of EEA1. This requirement is likely to underlie at least in part the essential role of Ca2+/CaM in endosome fusion. [References: 37]
机译：Ca2 +是膜融合中必不可少的要求，它通过结合蛋白（例如钙调蛋白（CaM））起作用。 Ca2 + / CaM是体外早期内体融合所必需的，但是，该要求的分子基础尚不清楚。内体融合的另一个要求是蛋白质早期内体抗原1（EEA1），其向内体的募集取决于磷脂酰肌醇3-磷酸[PI（3）P]和Rab5 GTPase。在本文中，我们证明了通过使用化学抑制剂或针对CaM的特异性抗体来抑制Ca2 + / CaM，会导致活细胞和体外对EEA1与内体膜结合的抑制作用都得到了深远的抑制。 EEA1从内体膜完全解离所需的Ca2 + / CaM抑制剂的浓度对磷脂酰肌醇3激酶的活性或内源性PI（3）P水平没有影响。但是，Ca2 + / CaM抑制剂可减少EEA1与含有PI（3）P的脂质体的相互作用。因此，Ca2 + / CaM似乎是EEA1与内体PI（3）P稳定相互作用所必需的，也许是通过直接或间接稳定EEA1的C端FYVE域的四级组织。该要求可能至少部分是Ca2 + / CaM在内体融合中的重要作用的基础。 [参考：37]

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