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The GxSxG motif of Arabidopsis monoacylglycerol lipase (MAGL6 and MAGL8) is essential for their enzyme activities

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Abstract

Monoacylglycerol lipase (MAGL) catalyzes the hydrolysis of monoacylglycerols (MAG) to free fatty acids and glycerol, which is the last step of triacylglycerol breakdown. Among sixteen members, Arabidopsis thaliana MAGL6 (AtMAGL6) and AtMAGL8 showed strong lipase activities, but several AtMAGLs including AtMAGL16 displayed very weak activities (Kim et al. in Plant. J 85:758–771, 2016). To understand the internal factors that influence Arabidopsis MAGL activities, this study investigated the significance of ‘GxSxS motif,’ which is conserved in MAGLs. First, we observed that the presence of a serine protease inhibitor, phenylmethylsulfonyl fluoride, decreased the enzyme activity of AtMAGL6 and AtMAGL8 by IC50 values of 2.30 and 2.35, respectively. Computational modeling showed that amino acid changes of the GxSxG motif in AtMAGL6 and AtMAGL8 altered the nucleophilic elbow structure, which is the active site of MAGLs. Mutating the GxSxG motif in the recombinant maltose binding protein (MBP):AtMAGL6 and MBP:AtMAGL8 proteins to SxSxG, GxAxG, and GxSxS motifs completely demolished the activities of the mutant MAGLs. In contrast, no significant differences were observed between the activities of AtMAGL16 wild type form harboring the SxSxG motif, and mutant AtMAGL16 containing the GxSxG motif. These results revealed that the glycine and serine residues of the GxSxG motif are essential for AtMAGL6 and AtMAGL8 enzyme activities, and that AtMAGL16 may not be involved in the hydrolysis of lipid substrates.

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Acknowledgments

This work was supported by grants from the National Research Foundation (NRF-2016R1A2B2010068) of Korea and the Next-Generation BioGreen 21 Program (No. PJ011052) of the Rural Development Administration, Republic of Korea.

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  1. Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, 61186, Republic of Korea

    Ryeo Jin Kim & Mi Chung Suh

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  1. Ryeo Jin Kim
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  2. Mi Chung Suh
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Correspondence to Mi Chung Suh.

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Kim, R.J., Suh, M.C. The GxSxG motif of Arabidopsis monoacylglycerol lipase (MAGL6 and MAGL8) is essential for their enzyme activities. Appl Biol Chem 59, 833–840 (2016). https://doi.org/10.1007/s13765-016-0232-1

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