{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T19:47:31Z","timestamp":1788292051325,"version":"build-2803163510"},"reference-count":27,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2017,3,26]],"date-time":"2017-03-26T00:00:00Z","timestamp":1490486400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Rare Trait Hope Fund"},{"name":"Vaincre l\u2019AGU"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Aspartylglucosaminidase (AGA) is a lysosomal hydrolase that participates in the breakdown of glycoproteins. 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 that contain either Ser or Thr in position 149 have been described, but it is unknown if this affects AGA processing or activity. Here, we have directly compared the Ser149\/Thr149 variants of AGA and show that they do not differ in terms of relative specific activity or processing. Therefore, Thr149 AGA, which is the rare variant, can be considered as a neutral or benign variant. Furthermore, we have here produced codon-optimized versions of these two variants and show that they are expressed at significantly higher levels than AGA with the natural codon-usage. Since optimal AGA expression is of vital importance for both gene therapy and enzyme replacement, our data suggest that use of codon-optimized AGA may be beneficial for these therapy options.<\/jats:p>","DOI":"10.3390\/ijms18040706","type":"journal-article","created":{"date-parts":[[2017,3,27]],"date-time":"2017-03-27T10:49:10Z","timestamp":1490611750000},"page":"706","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Functional Analysis of the Ser149\/Thr149 Variants of Human Aspartylglucosaminidase and Optimization of the Coding Sequence for Protein Production"],"prefix":"10.3390","volume":"18","author":[{"given":"Antje","family":"Banning","sequence":"first","affiliation":[{"name":"Institute of Biochemistry, Medical Faculty, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan F.","family":"K\u00f6nig","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry, Medical Faculty, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6240-8621","authenticated-orcid":false,"given":"Steven J.","family":"Gray","sequence":"additional","affiliation":[{"name":"Gene Therapy Center and Department of Ophthalmology, University of North Carolina, Chapel Hill, NC 27302, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8393-1825","authenticated-orcid":false,"given":"Ritva","family":"Tikkanen","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry, Medical Faculty, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,3,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1016\/0006-291X(66)90344-5","article-title":"Enzymatic cleavage of glycopeptides","volume":"24","author":"Makino","year":"1966","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1002\/j.1460-2075.1993.tb05656.x","article-title":"Lysosomal aspartylglucosaminidase is processed to the active subunit complex in the endoplasmic reticulum","volume":"12","author":"Ikonen","year":"1993","journal-title":"EMBO J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1102","DOI":"10.1038\/nsb1295-1102","article-title":"Three-dimensional structure of human lysosomal aspartylglucosaminidase","volume":"2","author":"Oinonen","year":"1995","journal-title":"Nat. Struct. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3210","DOI":"10.1074\/jbc.M210431200","article-title":"Two-step dimerization for autoproteolysis to activate glycosylasparaginase","volume":"278","author":"Wang","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1089\/dna.1995.14.305","article-title":"Intracellular sorting of aspartylglucosaminidase: The role of N-linked oligosaccharides and evidence of Man-6-P-independent lysosomal targeting","volume":"14","author":"Tikkanen","year":"1995","journal-title":"DNA Cell Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2954","DOI":"10.1002\/j.1460-2075.1996.tb00658.x","article-title":"Functional analyses of active site residues of human lysosomal aspartylglucosaminidase: Implications for catalytic mechanism and autocatalytic activation","volume":"15","author":"Tikkanen","year":"1996","journal-title":"EMBO J."},{"key":"ref_7","first-page":"1916","article-title":"Molecular genetics of aspartylglucosaminuria","volume":"107","author":"Palotie","year":"1991","journal-title":"Duodecim"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/S0140-6736(68)92355-6","article-title":"Aspartylglycosaminuria. An inborn error of metabolism associated with mental defect","volume":"2","author":"Pollitt","year":"1968","journal-title":"Lancet"},{"key":"ref_9","first-page":"1","article-title":"Aspartylglycosaminuria. Analysis of thirty-four patients","volume":"1","author":"Autio","year":"1972","journal-title":"J. Ment. Defic. Res."},{"key":"ref_10","first-page":"149","article-title":"Aspartylglycosaminuria (AGU). Further aspects on its clinical picture, mode of inheritance and epidemiology based on a series of 57 patients","volume":"5","author":"Autio","year":"1973","journal-title":"Ann. Clin. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1007\/BF00200902","article-title":"High prevalence of aspartylglycosaminuria among school-age children in eastern Finland","volume":"87","author":"Mononen","year":"1991","journal-title":"Hum. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1002\/j.1460-2075.1991.tb07920.x","article-title":"Aspartylglucosaminuria: cDNA encoding human aspartylglucosaminidase and the missense mutation causing the disease","volume":"10","author":"Ikonen","year":"1991","journal-title":"EMBO J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2941","DOI":"10.1073\/pnas.88.7.2941","article-title":"Aspartylglycosaminuria in the Finnish population: Identification of two point mutations in the heavy chain of glycoasparaginase","volume":"88","author":"Mononen","year":"1991","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1016\/0888-7543(91)90120-4","article-title":"In vitro mutagenesis helps to unravel the biological consequences of aspartylglucosaminuria mutation","volume":"11","author":"Ikonen","year":"1991","journal-title":"Genomics"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1002\/humu.1380050408","article-title":"Identification of a novel mutation causing aspartylglucosaminuria reveals a mutation hotspot region in the aspartylglucosaminidase gene","volume":"5","author":"Isoniemi","year":"1995","journal-title":"Hum. Mutat."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1002\/humu.1380010503","article-title":"Mutations causing aspartylglucosaminuria (AGU): A lysosomal accumulation disease","volume":"1","author":"Ikonen","year":"1992","journal-title":"Hum. Mutat."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1177\/0883073812469049","article-title":"Aspartylglucosaminuria: Unusual neonatal presentation in Qatari twins with a novel aspartylglucosaminidase gene mutation and 3 new cases in a Turkish family","volume":"29","author":"Opladen","year":"2014","journal-title":"J. Child. Neurol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1093\/hmg\/10.9.983","article-title":"Molecular pathogenesis of a disease: Structural consequences of aspartylglucosaminuria mutations","volume":"10","author":"Saarela","year":"2001","journal-title":"Hum. Mol. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"25320","DOI":"10.1074\/jbc.273.39.25320","article-title":"Activation and oligomerization of aspartylglucosaminidase","volume":"273","author":"Saarela","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"37583","DOI":"10.1038\/srep37583","article-title":"Identification of small molecule compounds for pharmacological chaperone therapy of aspartylglucosaminuria","volume":"6","author":"Banning","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1093\/hmg\/5.6.737","article-title":"Ser72Pro active-site disease mutation in human lysosomal aspartylglucosaminidase: Abnormal intracellular processing and evidence for extracellular activation","volume":"5","author":"Peltola","year":"1996","journal-title":"Hum. Mol. Genet."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1038\/nature15393","article-title":"A global reference for human genetic variation","volume":"526","author":"Auton","year":"2015","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1038\/nature15394","article-title":"An integrated map of structural variation in 2504 human genomes","volume":"526","author":"Sudmant","year":"2015","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/nature19057","article-title":"Analysis of protein-coding genetic variation in 60,706 humans","volume":"536","author":"Lek","year":"2016","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2073","DOI":"10.1093\/nar\/gks1205","article-title":"Sounds of silence: Synonymous nucleotides as a key to biological regulation and complexity","volume":"41","author":"Shabalina","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1016\/j.molmed.2014.09.003","article-title":"A critical analysis of codon optimization in human therapeutics","volume":"20","author":"Mauro","year":"2014","journal-title":"Trends Mol. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1089\/hum.2012.107","article-title":"Restoration of cytoskeleton homeostasis after gigaxonin gene transfer for giant axonal neuropathy","volume":"24","author":"Mussche","year":"2013","journal-title":"Hum. Gene Ther."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/4\/706\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:31:18Z","timestamp":1760207478000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/4\/706"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,3,26]]},"references-count":27,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2017,4]]}},"alternative-id":["ijms18040706"],"URL":"https:\/\/doi.org\/10.3390\/ijms18040706","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,3,26]]}}}