Neuferricin, a novel extracellular heme-binding protein, promotes neurogenesis
Neuferricin, a novel extracellular heme-binding protein, promotes neurogenesis
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DOI：
10.1111/j.1471-4159.2009.06522.x
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发表时间：
2010-03-01
影响因子：
4.7
通讯作者：
Fujimoto, Masafumi
中科院分区：
文献类型：
--
作者：
Kimura, Ikuo;Nakayama, Yoshiaki;Fujimoto, Masafumi
我们发现了一种新的细胞外血红素结合蛋白，并将其命名为neuferricin。在High Five细胞中产生的重组小鼠中铁蛋白被有效地分泌到培养基中。小鼠中性铁蛋白mRNA在胚胎期主要在大脑中表达，在发育过程中逐渐增加。在出生后，它广泛表达于大脑、心脏、肾上腺和肾脏。小鼠中性铁蛋白有263个氨基酸。它具有细胞色素b5样血红素/类固醇结合结构域，并且似乎与血红素结合，因为中性铁蛋白溶液，而不是中性铁蛋白Delta HBD溶液（缺乏血红素结合结构域的突变体），呈褐色，在402 nm处有吸光度峰。此外，利用血红素亲和层析法进行抗中性铁蛋白抗体实验，证实了在Neuro2a细胞培养基中检测到的内源性中性铁蛋白与血红素相关。在诱导Neuro2a细胞分化过程中，通过RNA干扰抑制内源性中性铁蛋白过度促进细胞存活和增殖，抑制神经突生长。添加重组小鼠中铁蛋白，而不添加中铁蛋白δ HBD，可以抑制Neuro2a细胞的存活，并使其免受中铁蛋白RNAi的影响。在原代培养的小鼠神经前体细胞中，重组小鼠中铁蛋白显示出促进神经发生的能力。neuferricin是一种具有细胞色素b5样血红素/类固醇结合域的新型细胞外血红素结合蛋白及其神经源性活性的鉴定，不仅为大脑发育提供了新的见解，而且为血红素结合蛋白作为细胞外信号传递体的功能提供了新的见解。
P>We identified a novel extracellular heme-binding protein and named it neuferricin. The recombinant mouse neuferricin produced in High Five cells was secreted efficiently into the culture medium. Mouse neuferricin mRNA was expressed mainly in the brain at the embryo stage and gradually increased during development. At postnatal stage, it was widely expressed in the brain, heart, adrenal gland, and kidney. Mouse neuferricin has 263 amino acids. It has a cytochrome b5-like heme/steroid-binding domain and appeared to bind hemin because neuferricin solution, but not a solution of neuferricin Delta HBD (a mutant lacking the heme-binding domain), was tinged with brown and had an absorbance peak at 402 nm. In addition, the experiment with anti-neuferricin antibody using heme-affinity chromatography proved that the endogenous neuferricin detected in the culture medium of Neuro2a cells was associated with hemin. Inhibition of endogenous neuferricin by RNA interference excessively promoted cell survival and proliferation and suppressed neurite outgrowth during the induction of differentiation in Neuro2a cells. Addition of recombinant mouse neuferricin, but not neuferricin Delta HBD, suppressed survival of Neuro2a cells and rescued from the effects of neuferricin RNAi. In primary cultured mouse neural precursor cells, recombinant mouse neuferricin exhibited the ability to promote neurogenesis. The identification of neuferricin, a novel extracellular heme-binding protein with cytochrome b5-like heme/steroid-binding domain and its neurogenic activity, provide new insights not only into brain development but also the function of heme-binding proteins as extracellular signal transmitters.