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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >FcεRI-mediated recruitment of p53/56~(lyn) to detergent-resistant membrane domains accompanies cellular signaling
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FcεRI-mediated recruitment of p53/56~(lyn) to detergent-resistant membrane domains accompanies cellular signaling

机译：FcεRI介导的p53 / 56〜（lyn）向去污剂抗性膜结构域募集伴随细胞信号转导

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摘要

Detergent-resistant plasma membrane structures, such as caveolae, have been implicated in signaling, transport, and vesicle trafficking functions. Using sucrose gradient ultracentrifugation, we have isolated low-density, Triton X-100-insoluble membrane domains from RBL-2H3 mucosal mast cells that contain several markers common to caveolae, including a src-family tyrosine kinase, pS3/56~(lyn). Aggregation of FceRI, the high-affinity IgE receptor, causes a significant increase in the amount of p53/56~(lyn) associated with these low-density membrane domains. Under our standard conditions for lysis, IgE-FcεRI fractionates with the majority of the solubilized proteins, whereas aggregated receptor complexes are found at a higher density in the gradient. Stimulated translocation of p53/56~(lyn) is accompanied by increased tyrosine phosphorylation of several proteins in the low-density membrane domains as well as enhanced in vitro tyrosine kinase activity toward these proteins and an exogenous substrate. With a lower detergent-to-ccll ratio during lysis, significant FceRI remains associated with these membrane domains, consistent with the ability to coimmu-noprecipitate tyrosine kinase activity with FceRI under similar lysis conditions [Pribluda, V. S., Pribluda, C. & Metzger, H. (1994) Proc. Natl. Acad. Sci. USA 91, 11246-11250]. These results indicate that specialized membrane domains may be directly involved in the coupling of receptor aggregation to the activation of signaling events.
机译：耐清洁剂的质膜结构（例如小窝）与信号传导，运输和囊泡运输功能有关。使用蔗糖梯度超速离心，我们从RBL-2H3粘膜肥大细胞中分离了低密度，Triton X-100不溶性膜结构域，该结构域包含海绵体常见的几种标志物，包括src家族酪氨酸激酶pS3 / 56〜（lyn） 。高亲和力IgE受体FceRI的聚集导致与这些低密度膜域相关的p53 / 56〜（lyn）数量显着增加。在我们的标准裂解条件下，IgE-FcεRI可与大多数可溶蛋白分馏，而聚集的受体复合物在梯度中的密度更高。 p53 / 56〜（lyn）的刺激易位伴随着低密度膜结构域中几种蛋白质酪氨酸磷酸化的增强，以及针对这些蛋白质和外源底物的体外酪氨酸激酶活性的增强。裂解期间较低的去垢剂/菌落比，显着的FceRI仍与这些膜结构域相关，这与在类似裂解条件下与FceRI共沉淀酪氨酸激酶活性的能力相一致[Pribluda，VS，Pribluda，C.＆Metzger， H.（1994），美国Natl。学院科学USA 91，11246-11250]。这些结果表明，专门的膜结构域可能直接参与受体聚集与信号转导事件的激活的耦合。

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