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Bioinformatic and biochemical studies point to AAGR-1 as the ortholog of human acid α-glucosidase in Caenorhabditis elegans

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Abstract

Human acid α-glucosidase (GAA, EC 3.2.1.20) is a lysosomal enzyme that belongs to the glycoside hydrolase family 31 (GH31) and catalyses the hydrolysis of α-1,4- and α-1,6-glucosidic linkages at acid pH. Hereditary deficiency of GAA results in lysosomal glycogen storage disease type II (GSDII, Pompe disease). The aim of this study was to assess GH31 proteins in Caenorhabditis elegans (C. elegans) to identify the ortholog of human GAA. Bioinformatic searches for GAA ortholog in C. elegans genome revealed four acid alpha-glucosidase-related (aagr-1–4) genes. Multiple sequence alignment of AAGRs with other GH31 proteins demonstrated their evolutionary conservation. Phylogenetic analyses suggested clustering of AAGR-1 and -2 with acid-active and AAGR-3 and -4 with neutral-active GH31 enzymes. In order to prove the AAGRs’ predicted α-glucosidase activity, we performed RNA interference of all four aagr genes. The impact on the α-glucosidase activity was evaluated at pH 4.0 (acid) and pH 6.5 (neutral), with or without the inhibitor acarbose. AAGR-1 and -2 expressed acidic α-glucosidase activity; on the contrary, AAGR-3 not -4 represented the predominant neutral α-glucosidase activity in C. elegans. Similar results were obtained in each of aagr-1 and -4 deletion mutants. Moreover, based on our structural models of AAGRs and these biochemical experiments, we hypothesize that the enzymatic sensitivity of AAGR-2 and human maltase-glucoamylase to the inhibitor acarbose is associated with a tyrosine residue in the GH31 active site, whereas acarbose resistance of AAGR-1 and human GAA is associated with the corresponding tryptophane in the active site. Acid-active AAGR-1 may thus represent the ortholog of human GAA in C. elegans.

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Acknowledgements

The authors would like to thank Dr. Karel Jelinek (Mathematical and Statistical Projects, Prague, Czech Republic) for initial homology modelling of AAGRs on the basis of YicI template. We would also like to thank Eliska Machalova for technical assistance. This study was funded by the research project 0021620806 from the Ministry of Education, Youth and Sports of the Czech Republic and, in part, by the grant 304/08/0970 from the Czech Science Foundation.

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  1. 1st Faculty of Medicine, Institute of Inherited Metabolic Disorders, Charles University in Prague, Ke Karlovu 2, Prague 2, Czech Republic

    Jakub Sikora, Jana Uřinovská, Filip Majer, Helena Poupětová, Jitka Hlavatá, Marta Kostrouchová, Jana Ledvinová & Martin Hřebíček

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  1. Jakub Sikora
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Correspondence to Jakub Sikora or Martin Hřebíček.

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The first two authors contributed equally to this work.

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Sikora, J., Uřinovská, J., Majer, F. et al. Bioinformatic and biochemical studies point to AAGR-1 as the ortholog of human acid α-glucosidase in Caenorhabditis elegans . Mol Cell Biochem 341, 51–63 (2010). https://doi.org/10.1007/s11010-010-0436-3

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