Literature summary for 3.5.1.26 extracted from

Banning, A.; Koenig, J.F.; Gray, S.J.; Tikkanen, R.
Functional analysis of the Ser149/Thr149 variants of human aspartylglucosaminidase and optimization of the coding sequence for protein production (2017), Int. J. Mol. Sci., 18, 706 .

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	processing and activation of aspartylglucosaminidase by autocatalytic cleavage, overview	Homo sapiens

Application

Application	Comment	Organism
medicine	use of codon-optimized AGA may be beneficial for the therapy options in treatment of the lysosomal storage disorder aspartylglucosaminuria (AGU)	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
gene AGA, DNA and amino acid sequence determination and analysis, genomic organization of Aurelia gene AGA and exon-intron structure, molecular phylogenetic analysis based on the nucleotide sequences of N-terminal nucleophile (Ntn)_asparaginase_2_like superfamily genes, quantitative real-time PCR enzyme expresssion analysis	Aurelia aurita
gene AGA, genotyping, recombinant expression of codon-optimized genes encoding enzyme variants S149 and T149 in HEK-293T and HeLa cells, the Thr149 variant exhibits a slightly higher expression level than the Ser149 variant. Recombinant expression of AGA variants in transgenic patient fibroblasts, both wild-type and AGUFin-major mutant type. The transfection with the AGA variants improves the lysosomal morphology in AGU fibroblasts, low transfection efficiency	Homo sapiens

Cloned (Comment)

Organism

gene AGA, DNA and amino acid sequence determination and analysis, genomic organization of Aurelia gene AGA and exon-intron structure, molecular phylogenetic analysis based on the nucleotide sequences of N-terminal nucleophile (Ntn)_asparaginase_2_like superfamily genes, quantitative real-time PCR enzyme expresssion analysis

Aurelia aurita

gene AGA, genotyping, recombinant expression of codon-optimized genes encoding enzyme variants S149 and T149 in HEK-293T and HeLa cells, the Thr149 variant exhibits a slightly higher expression level than the Ser149 variant. Recombinant expression of AGA variants in transgenic patient fibroblasts, both wild-type and AGUFin-major mutant type. The transfection with the AGA variants improves the lysosomal morphology in AGU fibroblasts, low transfection efficiency

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
additional information	single nucleotide polymorphism rs2228119 (NM_000027.3:c.446C>G - p.(Thr149Ser)) results in amino acid variation Ser vs. Thr at position 149 (base and amino acid variation in red) of the human AGA enzyme. Functional analysis of the Ser149/Thr149 variants of human aspartylglucosaminidase and optimization of the coding sequence for protein production. Codon-optimized versions of the two variants are expressed at significantly higher levels than AGA with the natural codon-usage. The second most common allele in Finland is a 2 bp deletion called AGUFin-minor (NM_000027.3; c.199_200del - p.Glu67fc*3). Genotype frequency of single nucleotide polymorphism (SNP) rs2228119 in various populations	Homo sapiens
R161Q/C163S	naturally occuring mutation, the AGUFin-major mutation is a combination of two missense mutations, abolishing a disulfide bond and destabilizing the AGA structure. The pathogenic C163S substitution is always combined with a functionally neutral Arg161Gln substitution. Mutations in the AGA gene result in aspartylglucosaminuria (AGU, OMIM 208400), a lysosomal storage disorder that is characterized by progressive loss of intellectual capabilities and some skeletal abnormalities. AGU patients are born seemingly normal, but the progressive course of the disease manifests in, e.g. developmental delay, loss of speech and coarse facial features early in childhood. In adulthood, most AGU patients are severely retarded and require special care	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism	Structure
bafilomycin A1	-	Aurelia aurita
Chloroquine	-	Aurelia aurita

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
lysosome	-	Homo sapiens	5764	-
lysosome	in the lysosomes, the C-termini of both alpha- and beta-subunits are trimmed by proteases, resulting in the mature form of AGA, though these trimming steps are not necessary for the catalytic activity of AGA	Aurelia aurita	5764	-
additional information	Aurelia AGA possesses an N-terminal signal peptide	Aurelia aurita	-	-
additional information	the pro-enzyme contains a 23 residues signal peptide	Homo sapiens	-	-

Organism

Organism	UniProt	Comment	Textmining
Aurelia aurita	-	clonal polyp strains established from adult female Aurelia jellyfish captured in the Seto Inland Sea	-
Homo sapiens	P20933	Finnish population	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	AGA contains two glycosylation sites at Asn38 and Asn308	Homo sapiens
glycoprotein	Aurelia AGA possesses potential N-glycosylation sites, Asn36, Asn48, Asn168, and Asn213	Aurelia aurita
proteolytic modification	in the lysosomes, the C-termini of both alpha- and beta-subunits are trimmed by proteases, resulting in the mature form of AGA, though these trimming steps are not necessary for the catalytic activity of AGA	Aurelia aurita
proteolytic modification	processing and activation of aspartylglucosaminidase by autocatalytic cleavage, overview. AGA is synthesized in the endoplasmic reticulum as a 346 amino acid (aa) polypeptide from which 23 residues of the signal peptide are removed. Very soon after synthesis in the endoplasmic reticulum, two AGA precursors homodimerize, inducing an autocatalytic cleavage of both precursors N-terminally to Thr206 into 27 kDa pro-alpha and 17 kDa subunits. After transport to lysosomes, the pro-alpha is C-terminally cleaved into 24 kDa mature alpha-subunit, whereas processing of the beta-subunit gives rise to the 14 kDa beta'-subunit. Neither of these lysosomal processing steps displays an effect on the enzyme activity	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
ephyra	moderate expression level	Aurelia aurita	-
additional information	AGA expression increases during strobilation, and is then decreased in medusae	Aurelia aurita	-
polyp	low expression level	Aurelia aurita	-
strobila	high expression level	Aurelia aurita	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	processing and activation of aspartylglucosaminidase by autocatalytic cleavage, overview	Homo sapiens	?	-	?

Subunits

Subunits	Comment	Organism
heterotetramer	(alphabeta)2	Aurelia aurita

Synonyms

Synonyms	Comment	Organism
AGA	-	Homo sapiens
AGA	-	Aurelia aurita
aspartylglucosaminidase	-	Homo sapiens
aspartylglucosaminidase	-	Aurelia aurita

General Information

General Information	Comment	Organism
evolution	belongs to the group of so-called N-terminal nucleophile (NTN) hydrolases. The members of the NTN hydrolase family, which in addition to AGA also include, e.g., the proteasome beta-subunit and penicillin acylase, show very little similarity at the amino acid sequence level, but they exhibit a highly similar folded structure	Homo sapiens
evolution	molecular phylogenetic analysis of aspartylglucosaminidases	Aurelia aurita
malfunction	defects in the AGA gene result in a lysosomal storage disorder, aspartylglucosaminuria (AGU), that manifests mainly as progressive mental retardation. A number of AGU missense mutations have been identified that result in reduced AGA activity. Human variants contain either Ser or Thr in position 149, and Thr149 AGA, which is the rare variant, can be considered as a neutral or benign variant. AGU mutations result in reduced AGA activity in patient cells. Mutations in the AGA gene result in aspartylglucosaminuria (AGU, OMIM 208400), a lysosomal storage disorder that is characterized by progressive loss of intellectual capabilities and some skeletal abnormalities. AGU patients are born seemingly normal, but the progressive course of the disease manifests in, e.g., developmental delay, loss of speech and coarse facial features early in childhood. In adulthood, most AGU patients are severely retarded and require special care	Homo sapiens
malfunction	lysosomal acidification inhibitors, chloroquine or bafilomycin A1, disturb medusa morphogenesis at the oral end, suggesting involvement of lysosomal hydrolases in strobilation	Aurelia aurita
additional information	AGA belongs to the group of so-called N-terminal nucleophile (NTN) hydrolases, as the free alpha-amino group of Thr206 is involved in the catalysis as the base, whereas the OH group of Thr206 functions as a nucleophile during the catalysis	Homo sapiens
physiological function	aspartylglucosaminidase (AGA) is a lysosomal hydrolase that participates in the breakdown of glycoproteins	Homo sapiens
physiological function	the life cycle of the moon jellyfish, Aurelia aurita, alternates between a benthic asexual polyp stage and a planktonic sexual medusa (jellyfish) stage. Transition from polyp to medusa is called strobilation. Aspartylglucosaminidase (AGA), a lysosomal hydrolase, is upregulated during strobilation	Aurelia aurita