The nuclear receptor-binding SET Domain (NSD) family consists of NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L1 histone methyltransferases that are crucial for chromatin remodeling. NSDs are implicated in developmental disorders such as Wolf-Hirschhorn and Sotos syndromes as well as various cancers including t(4; 14)(p16; q32) myeloma, an incurable cancer in plasma cells. NSDs have been the target of intensive study to understand their biological functions more fully and inform anti-cancer drug design. Recombinant protein expression and purification of human NSDs using an E. coli expression system are notoriously challenging, but the production of pure, stable, and active NSDs is essential for further studies. To overcome production challenges, we propose a cost-efficient approach optimized to produce a high yield of NSDs using a modified E. coli expression system. We found that tagging the NSDs with a human influenza hemagglutinin (HA) tag greatly improved the quality of the recombinant NSDs, resulting in more than 95% pure, stable, and active NSD-HAs, with an increase in production yield up to 22.4-fold and up to 6.25 mg/L from LB E. coli culture, and without further purification such as ion-exchange or size-exclusion chromatography.

中文翻译：

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带有人流感血凝素标签的致癌NSD1，NSD2和NSD3的高产量重组表达和纯化。

核受体结合SET域（NSD）家族由对染色质重塑至关重要的NSD1，NSD2 / MMSET / WHSC1和NSD3 / WHSC1L1组蛋白甲基转移酶组成。NSD与发展性疾病有关，例如Wolf-Hirschhorn和Sotos综合征以及各种癌症，包括t（4; 14）（p16; q32）骨髓瘤，这是浆细胞中无法治愈的癌症。NSD已成为深入研究的目标，以更充分地了解其生物学功能并为抗癌药物设计提供信息。众所周知，使用大肠杆菌表达系统重组蛋白表达和纯化人NSD具有挑战性，但是生产纯净，稳定和活跃的NSD对于进一步研究至关重要。为了克服生产挑战，我们提出了一种经济高效的方法，该方法经过优化，可使用改良的E生产高产量的NSD。大肠杆菌表达系统。我们发现，使用人流感血凝素（HA）标签标记NSD可以大大提高重组NSD的质量，从而产生超过95％的纯度，稳定和活性的NSD-HA，产量最多可提高22.4。可从LB大肠杆菌培养液中分离并纯化至最高6.25 mg / L，无需进一步纯化（例如离子交换色谱法或尺寸排阻色谱法）。