With many crucial roles in enzymatic aerobic metabolism, the concentration of the heme must be tightly regulated. The heme exporter Feline Leukemia Virus sub-group C Receptor 1a (FLVCR1a), an integral membrane protein with twelve transmembrane helices, is a key player in the maintenance of cellular heme homeostasis. It was first identified as the host receptor for the Feline Leukemia Virus sub-group C (FeLV-C), a retrovirus causing hematological abnormalities in cats and other felines. Mutations in the Flvcr1 were later identified in human patients affected by Posterior Column Ataxia and Retinitis Pigmentosa (PCARP) and Hereditary Sensory and Autonomic Neuropathies (HSANs). Despite being an essential component in heme balance, currently there is a lack in the understanding of its function at the molecular level, including the effect of disease-causing mutations on protein function and structure. Therefore, there is a need for protocols to achieve efficient recombinant production yielding milligram amounts of highly pure protein to be used for biochemical and structural studies. Here, we report the first FLVCR1a reliable protocol suitable for both antibody generation and structural characterisation.

中文翻译：

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血红素输出蛋白FLVCR1a的表达和纯化。

血红素的浓度在酶促有氧代谢中起着至关重要的作用，必须严格调节血红素的浓度。血红素出口商猫白血病病毒C亚型受体1a（FLVCR1a）是具有十二个跨膜螺旋的完整膜蛋白，是维持细胞血红素稳态的关键因素。它首先被确定为猫白血病病毒C亚组（FeLV-C）的宿主受体，后者是一种引起猫和其他猫科血液学异常的逆转录病毒。Flvcr1的突变后来在受后柱共济失调和色素性视网膜炎（PCARP）和遗传性感觉和自主神经病（HSAN）影响的人类患者中被发现。尽管它是血红素平衡的重要组成部分，但目前在分子水平尚缺乏对其功能的了解，包括致病突变对蛋白质功能和结构的影响。因此，需要一种协议以实现有效的重组生产，从而产生毫克量的用于生物化学和结构研究的高纯度蛋白质。在这里，我们报告了适用于抗体生成和结构表征的第一个FLVCR1a可靠方案。