Effect of Erythropoietin, Iron Deficiency and Iron Overload on Liver Matriptase-2 (TMPRSS6) Protein Content in Mice and Rats.
Effect of Erythropoietin, Iron Deficiency and Iron Overload on Liver Matriptase-2 (TMPRSS6) Protein Content in Mice and Rats.
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DOI：
10.1371/journal.pone.0148540
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Krijt J
中科院分区：
文献类型：
--
作者：
Frýdlová J;Přikryl P;Truksa J;Falke LL;Du X;Gurieva I;Vokurka M;Krijt J
间质蛋白酶-2（TMPRSS 6）是铁调素表达的重要负调节因子;然而，体内铁过载或加速红细胞生成对肝脏TMPRSS 6蛋白含量的影响在很大程度上是未知的。我们确定TMPRSS 6蛋白质含量的质膜富集肝匀浆的馏分，通过免疫印迹，使用商业抗体提出了对TMPRSS 6的催化结构域。通过在3000 g下离心并洗涤获得质膜富集级分。在小鼠和大鼠中，在3000 μ g级分中检测到TMPRSS 6作为120 kDa全长蛋白。喂养缺铁饮食以及促红细胞生成素治疗增加TMPRSS 6蛋白含量的大鼠和小鼠的转录后机制，促红细胞生成素增加TMPRSS 6蛋白也观察到Bmp 6突变小鼠。向小鼠施用高剂量铁（200、350和700 mg/kg）降低TMPRSS 6蛋白质含量。在对照动物的质膜富集组分中检测到Hemojuvelin，其为约52 kDa的全长蛋白质;在缺铁动物中，全长蛋白质在N-末端部分裂解，产生约47 kDa的额外弱条带。在从hemojuvelin突变小鼠的肝脏中，TMPRSS 6蛋白质含量强烈下降，表明完整的hemojuvelin是必要的TMPRSS 6在膜中的稳定表达。总体而言，结果表明铁状态和促红细胞生成素管理的肝脏TMPRSS 6蛋白的转录后调节，并提供支持TMPRSS 6和hemojuvelin蛋白在体内的相互作用。
Matriptase-2 (TMPRSS6) is an important negative regulator of hepcidin expression; however, the effects of iron overload or accelerated erythropoiesis on liver TMPRSS6 protein content in vivo are largely unknown. We determined TMPRSS6 protein content in plasma membrane-enriched fractions of liver homogenates by immunoblotting, using a commercial antibody raised against the catalytic domain of TMPRSS6. Plasma membrane-enriched fractions were obtained by centrifugation at 3000 g and washing. TMPRSS6 was detected in the 3000 g fraction as a 120 kDa full-length protein in both mice and rats. Feeding of iron-deficient diet as well as erythropoietin treatment increased TMPRSS6 protein content in rats and mice by a posttranscriptional mechanism; the increase in TMPRSS6 protein by erythropoietin was also observed in Bmp6-mutant mice. Administration of high doses of iron to mice (200, 350 and 700 mg/kg) decreased TMPRSS6 protein content. Hemojuvelin was detected in the plasma membrane-enriched fractions of control animals as a full length protein of approximately 52 kDa; in iron deficient animals, the full length protein was partially cleaved at the N-terminus, resulting in an additional weak band of approximately 47 kDa. In livers from hemojuvelin-mutant mice, TMPRSS6 protein content was strongly decreased, suggesting that intact hemojuvelin is necessary for stable TMPRSS6 expression in the membrane. Overall, the results demonstrate posttranscriptional regulation of liver TMPRSS6 protein by iron status and erythropoietin administration, and provide support for the interaction of TMPRSS6 and hemojuvelin proteins in vivo.