FC-EPSILON-RI-MEDIATED RECRUITMENT OF P53/56(LYN) TO DETERGENT-RESISTANT MEMBRANE DOMAINS ACCOMPANIES CELLULAR SIGNALING
FC-EPSILON-RI-MEDIATED RECRUITMENT OF P53/56(LYN) TO DETERGENT-RESISTANT MEMBRANE DOMAINS ACCOMPANIES CELLULAR SIGNALING
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DOI：
10.1073/pnas.92.20.9201
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发表时间：
1995-09-26
影响因子：
11.1
通讯作者：
BAIRD, B
中科院分区：
文献类型：
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作者：
FIELD, KA;HOLOWKA, D;BAIRD, B
抗洗涤剂的质膜结构，例如小窝，与信号传导、转运和囊泡运输功能有关。使用蔗糖梯度超速离心，我们从RBL-2 H3粘膜肥大细胞中分离出低密度、Triton X-100不溶性膜结构域，这些细胞含有小窝常见的几种标记物，包括SRC家族酪氨酸激酶p53/56（林恩）。高亲和力IgE受体Fc ε RI的聚集导致与这些低密度膜结构域相关的p53/56（林恩）的量显著增加。在我们的标准裂解条件下，IgE-Fc β RI与大部分溶解的蛋白质分离，而聚集的受体复合物在梯度中以较高的密度存在。p53/56（林恩）的刺激易位伴随着低密度膜结构域中几种蛋白质的酪氨酸磷酸化增加以及对这些蛋白质和外源性底物的体外酪氨酸激酶活性增强。在裂解过程中洗涤剂与细胞的比率较低的情况下，显著的Fc β RI保持与这些膜结构域相关，这与在类似的裂解条件下酪氨酸激酶活性与Fc γ RI免疫共沉淀的能力一致[Pribluda，V，S.，普里布卢达角gr Metzger，H.，（1994）Proc. Natl. Acad. Sci，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的耦合信号事件的激活。
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, p53/56(lyn). Aggregation of Fc epsilon RI, 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-Fc epsilon RI 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-cell ratio during lysis, significant Fc epsilon RI remains associated with these membrane domains, consistent with the ability to coimmunoprecipitate tyrosine kinase activity with Fc epsilon RI under similar lysis conditions [Pribluda, V, S., Pribluda, C. gr 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.