Structural and functional analysis of plant ELO-like elongase for fatty acid elongation
Date
2024-08
Authors
Meesapyodsuk, Dauenpen
Sun, Kaiwen
Qiu, Xiao
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Plant Molecular Biology
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Article
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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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This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s11103-024-01490-5
Keywords
Elongase, Eranthis hyemalis, site-directed mutagenesis, structure-function, 3-ketoacyl-CoA synthase, very long chain polyunsaturated fatty acids
Citation
Meesapyodsuk, D., Sun, K., & Qiu, X. (2024). Structural and functional analysis of plant ELO-like elongase for fatty acid elongation. Plant molecular biology, 114(5), 90. https://doi.org/10.1007/s11103-024-01490-5
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DOI
https://doi.org/10.1007/s11103-024-01490-5