Fortilin interacts with TGF-β1 and prevents TGF-β receptor activation.
Fortilin interacts with TGF-β1 and prevents TGF-β receptor activation.
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DOI：
10.1038/s42003-022-03112-6
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发表时间：
2022-02-23
影响因子：
5.9
通讯作者：
Fujise K
中科院分区：
文献类型：
--
作者：
Pinkaew D;Martinez-Hackert E;Jia W;King MD;Miao F;Enger NR;Silakit R;Ramana K;Chen SY;Fujise K
Fortilin是一种由172个氨基酸组成的多功能蛋白质，存在于细胞内外。虽然fortilin结合和调节各种细胞蛋白，细胞外fortilin的生物学作用仍然未知。在这里，我们报告fortilin特异性地与TGF-β1相互作用，并阻止其激活TGF-β1信号通路。在标准免疫沉淀-蛋白质印迹分析中，福替林共免疫沉淀TGF-β1及其亚型。改良的ELISA法显示，人血清中TGF-β1与Fortilin保持复合。生物层干涉法和表面等离子体共振（SPR）均表明Fortilin与TGF-β1直接结合。SPR分析还表明，Fortilin和TGF-β受体II（TGFβRII）竞争TGF-β1。荧光素酶和分泌型碱性磷酸酶报告基因检测均显示fortilin抑制TGF-β1激活Smad 3与Smad结合元件的结合。Fortilin在定量蛋白质印迹分析和ELISA中均抑制Smad 3的磷酸化。最后，Fortilin抑制TGFβ-1诱导的C3 H10 T1/2间充质祖细胞向平滑肌细胞的分化。计算机辅助虚拟对接显示，Fortilin占据了TGF-β1的口袋，而TGF-β1通常被TGFβRII占据，并且TGF-β1可以在任何给定时间结合Fortilin或TGFβRII。这些数据支持细胞外fortilin作为TGF-β1信号通路的负调节剂的作用。Fortilin通过占据TGF-β1的口袋并与TGF-βRII竞争结合TGF-β1来阻止TGF-β1受体的活化。这抑制了Smad 3磷酸化和C3 H10 T1/2间充质祖细胞向平滑肌细胞的分化。
Fortilin is a 172-amino acid multifunctional protein present in both intra- and extracellular spaces. Although fortilin binds and regulates various cellular proteins, the biological role of extracellular fortilin remains unknown. Here we report that fortilin specifically interacts with TGF-β1 and prevents it from activating the TGF-β1 signaling pathway. In a standard immunoprecipitation-western blot assay, fortilin co-immunoprecipitates TGF-β1 and its isoforms. The modified ELISA assay shows that TGF-β1 remains complexed with fortilin in human serum. Both bio-layer interferometry and surface plasmon resonance (SPR) reveal that fortilin directly bind TGF-β1. The SPR analysis also reveals that fortilin and the TGF-β receptor II (TGFβRII) compete for TGF-β1. Both luciferase and secreted alkaline phosphatase reporter assays show that fortilin prevents TGF-β1 from activating Smad3 binding to Smad-binding element. Fortilin inhibits the phosphorylation of Smad3 in both quantitative western blot assays and ELISA. Finally, fortilin inhibits TGFβ-1-induced differentiation of C3H10T1/2 mesenchymal progenitor cells to smooth muscle cells. A computer-assisted virtual docking reveals that fortilin occupies the pocket of TGF-β1 that is normally occupied by TGFβRII and that TGF-β1 can bind either fortilin or TGFβRII at any given time. These data support the role of extracellular fortilin as a negative regulator of the TGF-β1 signaling pathway. Fortilin prevents the activation of the TGF-β1 receptor by occupying the pocket of TGF-β1 and competing with TGF-βRII to bind with TGF-β1. This inhibits Smad3 phosphorylation and the differentiation of C3H10T1/2 mesenchymal progenitor cells to smooth muscle cells.
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DOI：
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发表时间：
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