Structural and functional analysis of plant ELO-like elongase for fatty acid elongation.

IF 3.9 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Plant Molecular Biology Pub Date : 2024-08-22 DOI:10.1007/s11103-024-01490-5
Dauenpen Meesapyodsuk, Kaiwen Sun, Xiao Qiu
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Abstract

ELO-like elongase is a condensing enzyme elongating long chain fatty acids in eukaryotes. Eranthis hyemalis ELO-like elongase (EhELO1) is the first higher plant ELO-type elongase that is highly active in elongating a wide range of polyunsaturated fatty acids (PUFAs) and some monounsaturated fatty acids (MUFAs). This study attempted using domain swapping and site-directed mutagenesis of EhELO1 and EhELO2, a close homologue of EhELO1 but with no apparent elongase activity, to elucidate the structural determinants critical for catalytic activity and substrate specificity. Domain swapping analysis of the two showed that subdomain B in the C-terminal half of EhELO1 is essential for MUFA elongation while subdomain C in the C-terminal half of EhELO1 is essential for both PUFA and MUFA elongations, implying these regions are critical in defining the architecture of the substrate tunnel for substrate specificity. Site-directed mutagenesis showed that the glycine at position 220 in the subdomain C plays a key role in differentiating the function of the two elongases. In addition, valine at 161 and cysteine at 165 in subdomain A also play critical roles in defining the architecture of the deep substrate tunnel, thereby contributing significantly to the acceptance of, and interaction with primer substrates.

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用于脂肪酸伸长的植物 ELO 样伸长酶的结构和功能分析。
ELO 样伸长酶是真核生物中伸长长链脂肪酸的一种缩合酶。Eranthis hyemalis ELO 样伸长酶(EhELO1)是第一个高等植物 ELO 型伸长酶,在伸长多种多不饱和脂肪酸(PUFA)和一些单不饱和脂肪酸(MUFA)方面具有很高的活性。本研究试图通过对 EhELO1 和 EhELO2(EhELO1 的近源同源物,但没有明显的伸长酶活性)进行结构域交换和定点突变,来阐明对催化活性和底物特异性至关重要的结构决定因素。对二者进行的结构域交换分析表明,EhELO1 C端半部分的子域B对MUFA的伸长至关重要,而EhELO1 C端半部分的子域C对PUFA和MUFA的伸长都至关重要,这意味着这些区域对底物特异性底物隧道结构的确定至关重要。定点突变显示,亚域 C 中 220 位的甘氨酸在区分两种伸长酶的功能方面起着关键作用。此外,亚域 A 中 161 位的缬氨酸和 165 位的半胱氨酸在确定深层底物隧道的结构方面也起着关键作用,从而对底物的接受和与底物的相互作用做出了重要贡献。
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来源期刊
Plant Molecular Biology
Plant Molecular Biology 生物-生化与分子生物学
自引率
2.00%
发文量
95
审稿时长
1.4 months
期刊介绍: Plant Molecular Biology is an international journal dedicated to rapid publication of original research articles in all areas of plant biology.The Editorial Board welcomes full-length manuscripts that address important biological problems of broad interest, including research in comparative genomics, functional genomics, proteomics, bioinformatics, computational biology, biochemical and regulatory networks, and biotechnology. Because space in the journal is limited, however, preference is given to publication of results that provide significant new insights into biological problems and that advance the understanding of structure, function, mechanisms, or regulation. Authors must ensure that results are of high quality and that manuscripts are written for a broad plant science audience.
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