Association of type 3 protein kinase C with focal contacts in rat embryo fibroblasts.
Association of type 3 protein kinase C with focal contacts in rat embryo fibroblasts.
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DOI：
10.1083/jcb.109.2.697
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发表时间：
1989-08
期刊：
影响因子：
--
通讯作者：
Klauck T
中科院分区：
文献类型：
--
作者：
Jaken S;Leach K;Klauck T
我们用免疫细胞荧光技术测定了Ca ~（2+）、磷脂依赖性蛋白激酶、蛋白激酶C（PKC）的亚细胞分布。使用对3型（α）PKC特异的单克隆抗体，我们已经确定在正常大鼠胚胎成纤维细胞（REF 52细胞）中至少有两种PKC池：弥漫性胞质和纤维相关。在固定和染色之前用螯合剂和去污剂提取除去细胞质PKC。纤维相关染色保留在这些细胞骨架制剂中。细胞骨架3型PKC染色与黏着斑蛋白的染色非常相似，并与另一种黏着斑蛋白talin共定位。细胞松弛素，而不是秋水仙素，协调破坏粘着斑蛋白和PKC的染色模式。通过用佛波醇酯处理激活PKC引起微丝解聚和黏着斑蛋白染色的重组。我们建议，3型蛋白激酶C是一个调制组件的焦点接触，并有一个主要的作用，在调节协会的微丝束与质膜。
We have used immunocytofluorescence techniques to determine the subcellular distribution of the Ca2+, phospholipid-dependent protein kinase, protein kinase C (PKC). Using monoclonal antibodies that are specific for Type 3 (alpha) PKC, we have determined that there are least two pools of PKC in normal rat embryo fibroblasts (REF52 cells): diffuse cytoplasmic and fiber-associated. Extraction with chelators and detergent before fixing and staining removes the cytoplasmic PKC. The fiber-associated staining remains in these cytoskeleton preparations. The cytoskeleton Type 3 PKC staining closely resembles that of the focal contact protein vinculin and colocalizes with another focal contact protein, talin. Cytochalasin, but not colchicine, coordinately disrupts the staining pattern of vinculin and PKC. Activation of PKC by treatment with phorbol esters causes depolymerization of microfilaments and reorganization of vinculin staining. We propose that Type 3 PKC is a modulatory component of the focal contact and has a primary role in regulation of the association of microfilament bundles with the plasma membrane.