NLIP and HAD-like Domains of Pah1 and Lipin 1 Phosphatidate Phosphatases Are Essential for Their Catalytic Activities.

NLIP and HAD-like Domains of Pah1 and Lipin 1 Phosphatidate Phosphatases Are Essential for Their Catalytic Activities.
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DOI:
10.3390/molecules26185470
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发表时间:
2021-09-08
期刊:
Molecules (Basel, Switzerland)
影响因子:
--
通讯作者:
Hsieh LS
Hsieh LS
中科院分区:
其他
文献类型:
--
作者:
Hsu WH;Huang YH;Chen PR;Hsieh LS

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酿酒酵母Pah1磷酸酯磷酸酶(PAP)催化磷酸酯去磷酸化生成二酰基甘油,控制磷脂和三酰基甘油代谢。Pah1和human Lipin 1是内在无序蛋白,分别有56%和43%的未折叠区域。保守区和非保守区截断分析表明,N-和c -保守区对Pah1的催化活性至关重要。在pah和Lipin 1的保守的n端Lipin (NLIP)结构域和c端Lipin (CLIP)/卤酸脱卤酶(HAD)样结构域中可以检测到PAP活性,这表明进化上保守的结构域对催化活性至关重要。无序亲水区域的去除大大降低了Pah1的蛋白质溶解度。硫氧还蛋白是一种在大肠杆菌中生产可溶性NLIP-HAD重组蛋白的高效融合蛋白。
Saccharomyces cerevisiae Pah1 phosphatidate phosphatase (PAP) catalyzes the dephosphorylation of phosphatidate to yield diacylglycerol, controlling phospholipids and triacylglycerol metabolisms. Pah1 and human Lipin 1 are intrinsically disordered proteins with 56% and 43% unfolded regions, respectively. Truncation analysis of the conserved and non-conserved regions showed that N- and C-conserved regions are essential for the catalytic activity of Pah1. PAP activities can be detected in the conserved N-terminal Lipin (NLIP) domain and C-terminal Lipin (CLIP)/haloacid dehalogenase (HAD)-like domain of Pah1 and Lipin 1, suggesting that the evolutionarily conserved domains are essential for the catalytic activity. The removal of disordered hydrophilic regions drastically reduced the protein solubility of Pah1. Thioredoxin is an efficient fusion protein for production of soluble NLIP–HAD recombinant proteins in Escherichia coli.
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