title: Autoproteolytic activation of human aspartylglucosaminidase.
abstract: Aspartylglucosaminidase (AGA) belongs to the N-terminal nucleophile (Ntn) hydrolase superfamily characterized by an N-terminal nucleophile as the catalytic residue. Three-dimensional structures of the Ntn hydrolases reveal a common folding pattern and equivalent stereochemistry at the active site. The activation of the precursor polypeptide occurs autocatalytically, and for some amidohydrolases of prokaryotes, the precursor structure is known and activation mechanisms are suggested. In humans, the deficient AGA activity results in a lysosomal storage disease, aspartylglucosaminuria (AGU) resulting in progressive neurodegeneration. Most of the disease-causing mutations lead to defective molecular maturation of AGA, and, to understand the structure-function relationship better, in the present study, we have analysed the effects of targeted amino acid substitutions on the activation process of human AGA. We have evaluated the effect of the previously published mutations and, in addition, nine novel mutations were generated. We could identify one novel amino acid, Gly258, with an important structural role on the autocatalytic activation of human AGA, and present the molecular mechanism for the autoproteolytic activation of the eukaryotic enzyme. Based on the results of the present study, and by comparing the available information on the activation of the Ntn-hydrolases, the autocatalytic processes of the prokaryotic and eukaryotic enzymes share common features. First, the critical nucleophile functions both as the catalytic and autocatalytic residue; secondly, the side chain of this nucleophile is oriented towards the scissile peptide bond; thirdly, conformational strain exists in the precursor at the cleavage site; finally, water molecules are utilized in the activation process.
authors: Department of Medical Genetics and National Public Health Institute, University of Helsinki, Haartmaninkatu 8, FIN-00290 Helsinki, Finland.
authors: Jani
authors: J
authors: Saarela
authors: Carita
authors: C
authors: Oinonen
authors: Anu
authors: A
authors: Jalanko
authors: Juha
authors: J
authors: Rouvinen
authors: Leena
authors: L
authors: Peltonen
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001227:Aspartylglucosylaminase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D002384:Catalysis; D002522:Chlorocebus aethiops; D004789:Enzyme Activation; D006801:Humans; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D013912:Threonine / Q000737:chemistry
chemical_list: D000596:Amino Acids; D013912:Threonine; D001227:Aspartylglucosylaminase