{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T14:26:04Z","timestamp":1791815164359,"version":"4.3.4"},"reference-count":35,"publisher":"National Academy of Sciences","issue":"36","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2013,9,3]]},"abstract":"<jats:p>\n                    <jats:italic>N<\/jats:italic>\n                    -glycosylation is a major posttranslational modification that endows proteins with various functions. It is established that\n                    <jats:italic>N<\/jats:italic>\n                    -glycans are essential for the correct folding and stability of some enzymes; however, the actual effects of\n                    <jats:italic>N<\/jats:italic>\n                    -glycans on their activities are poorly understood. Here, we show that human \u03b1-\n                    <jats:sc>l<\/jats:sc>\n                    -iduronidase (hIDUA), of which a dysfunction causes accumulation of dermatan\/heparan sulfate leading to mucopolysaccharidosis type I, uses its own\n                    <jats:italic>N<\/jats:italic>\n                    -glycan as a substrate binding and catalytic module. Structural analysis revealed that the mannose residue of the\n                    <jats:italic>N<\/jats:italic>\n                    -glycan attached to N372 constituted a part of the substrate-binding pocket and interacted directly with a substrate. A deglycosylation study showed that enzyme activity was highly correlated with the\n                    <jats:italic>N<\/jats:italic>\n                    -glycan attached to N372. The kinetics of native and deglycosylated hIDUA suggested that the\n                    <jats:italic>N<\/jats:italic>\n                    -glycan is also involved in catalytic processes. Our study demonstrates a previously unrecognized function of\n                    <jats:italic>N<\/jats:italic>\n                    -glycans.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1306939110","type":"journal-article","created":{"date-parts":[[2013,8,19]],"date-time":"2013-08-19T21:59:24Z","timestamp":1376949564000},"page":"14628-14633","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":42,"title":["Human \u03b1-\n                    <scp>l<\/scp>\n                    -iduronidase uses its own\n                    <i>N<\/i>\n                    -glycan as a substrate-binding and catalytic module"],"prefix":"10.1073","volume":"110","author":[{"given":"Nobuo","family":"Maita","sequence":"first","affiliation":[{"name":"Laboratory of X-Ray Crystallography, Institute for Enzyme Research, University of Tokushima, Tokushima 770-8503, Japan;"},{"name":"Division of Disease Proteomics, Institute for Enzyme Research, University of Tokushima, Tokushima 770-8503, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiro","family":"Tsukimura","sequence":"additional","affiliation":[{"name":"Department of Functional Bioanalysis, Meiji Pharmaceutical University, Tokyo 204-8588, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takako","family":"Taniguchi","sequence":"additional","affiliation":[{"name":"Division of Disease Proteomics, Institute for Enzyme Research, University of Tokushima, Tokushima 770-8503, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seiji","family":"Saito","sequence":"additional","affiliation":[{"name":"Department of Medical Management and Informatics, Hokkaido Information University, Hokkaido 069-8585, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuki","family":"Ohno","sequence":"additional","affiliation":[{"name":"Research Section, NPO for the Promotion of Research on Intellectual Property Tokyo, Tokyo 100-0005, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hisaaki","family":"Taniguchi","sequence":"additional","affiliation":[{"name":"Division of Disease Proteomics, Institute for Enzyme Research, University of Tokushima, Tokushima 770-8503, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hitoshi","family":"Sakuraba","sequence":"additional","affiliation":[{"name":"Department of Analytical Biochemistry, Meiji Pharmaceutical University, Tokyo 204-8588, Japan; and"},{"name":"Department of Clinical Genetics, Meiji Pharmaceutical University, Tokyo 204-8588, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2013,8,19]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"crossref","first-page":"2364","DOI":"10.1126\/science.291.5512.2364","article-title":"Intracellular functions of N-linked glycans","volume":"291","author":"Helenius A","year":"2001","unstructured":"A Helenius, M Aebi, Intracellular functions of N-linked glycans. Science 291, 2364\u20132369 (2001).","journal-title":"Science"},{"key":"e_1_3_4_2_2","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1016\/j.bmc.2009.02.037","article-title":"The effect of individual N-glycans on enzyme activity","volume":"17","author":"Skropeta D","year":"2009","unstructured":"D Skropeta, The effect of individual N-glycans on enzyme activity. Bioorg Med Chem 17, 2645\u20132653 (2009).","journal-title":"Bioorg Med Chem"},{"key":"e_1_3_4_3_2","doi-asserted-by":"crossref","first-page":"4411","DOI":"10.1021\/bi200346n","article-title":"The expanding horizons of asparagine-linked glycosylation","volume":"50","author":"Larkin A","year":"2011","unstructured":"A Larkin, B Imperiali, The expanding horizons of asparagine-linked glycosylation. Biochemistry 50, 4411\u20134426 (2011).","journal-title":"Biochemistry"},{"key":"e_1_3_4_4_2","doi-asserted-by":"crossref","first-page":"14861","DOI":"10.1016\/S0021-9258(18)82412-7","article-title":"Human acid \u03b2-glucosidase. N-glycosylation site occupancy and the effect of glycosylation on enzymatic activity","volume":"268","author":"Berg-Fussman A","year":"1993","unstructured":"A Berg-Fussman, ME Grace, YA Ioannou, GA Grabowski, Human acid \u03b2-glucosidase. N-glycosylation site occupancy and the effect of glycosylation on enzymatic activity. J Biol Chem 268, 14861\u201314866 (1993).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_5_2","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1042\/bj3320789","article-title":"Human \u03b1-galactosidase A: Glycosylation site 3 is essential for enzyme solubility","volume":"332","author":"Ioannou YA","year":"1998","unstructured":"YA Ioannou, KM Zeidner, ME Grace, RJ Desnick, Human \u03b1-galactosidase A: Glycosylation site 3 is essential for enzyme solubility. Biochem J 332, 789\u2013797 (1998).","journal-title":"Biochem J"},{"key":"e_1_3_4_6_2","doi-asserted-by":"crossref","first-page":"1591","DOI":"10.1002\/humu.21111","article-title":"Structural aspects of therapeutic enzymes to treat metabolic disorders","volume":"30","author":"Kang TS","year":"2009","unstructured":"TS Kang, RC Stevens, Structural aspects of therapeutic enzymes to treat metabolic disorders. Hum Mutat 30, 1591\u20131610 (2009).","journal-title":"Hum Mutat"},{"key":"e_1_3_4_7_2","first-page":"3421","volume-title":"The Metabolic and Molecular Bases of Inherited Disease","author":"Neufeld EF","year":"2001","unstructured":"EF Neufeld, J Muenzer The Metabolic and Molecular Bases of Inherited Disease, eds CR Scriver, AL Beaudet, WS Sly, D Valle (McGraw-Hill, 8th Ed, New York), pp. 3421\u20133452 (2001).","edition":"8"},{"key":"e_1_3_4_8_2","doi-asserted-by":"crossref","first-page":"22758","DOI":"10.1074\/jbc.272.36.22758","article-title":"Carbohydrate structures of recombinant human \u03b1-L-iduronidase secreted by Chinese hamster ovary cells","volume":"272","author":"Zhao KW","year":"1997","unstructured":"KW Zhao, KF Faull, ED Kakkis, EF Neufeld, Carbohydrate structures of recombinant human \u03b1-L-iduronidase secreted by Chinese hamster ovary cells. J Biol Chem 272, 22758\u201322765 (1997).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_9_2","doi-asserted-by":"crossref","first-page":"9695","DOI":"10.1073\/pnas.88.21.9695","article-title":"Human \u03b1-L-iduronidase: cDNA isolation and expression","volume":"88","author":"Scott HS","year":"1991","unstructured":"HS Scott, et al., Human \u03b1-L-iduronidase: cDNA isolation and expression. Proc Natl Acad Sci USA 88, 9695\u20139699 (1991).","journal-title":"Proc Natl Acad Sci USA"},{"key":"e_1_3_4_10_2","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1017\/S1462399408000550","article-title":"The mucopolysaccharidoses: A success of molecular medicine","volume":"10","author":"Clarke LA","year":"2008","unstructured":"LA Clarke, The mucopolysaccharidoses: A success of molecular medicine. Expert Rev Mol Med 10, e1 (2008).","journal-title":"Expert Rev Mol Med"},{"key":"e_1_3_4_11_2","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/1750-1172-3-24","article-title":"The prevalence of and survival in Mucopolysaccharidosis I: Hurler, Hurler-Scheie and Scheie syndromes in the UK","volume":"3","author":"Moore D","year":"2008","unstructured":"D Moore, MJ Connock, E Wraith, C Lavery, The prevalence of and survival in Mucopolysaccharidosis I: Hurler, Hurler-Scheie and Scheie syndromes in the UK. Orphanet J Rare Dis 3, 24 (2008).","journal-title":"Orphanet J Rare Dis"},{"key":"e_1_3_4_12_2","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1056\/NEJM200101183440304","article-title":"Enzyme-replacement therapy in mucopolysaccharidosis I","volume":"344","author":"Kakkis ED","year":"2001","unstructured":"ED Kakkis, et al., Enzyme-replacement therapy in mucopolysaccharidosis I. N Engl J Med 344, 182\u2013188 (2001).","journal-title":"N Engl J Med"},{"key":"e_1_3_4_13_2","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1016\/S0959-440X(97)80072-3","article-title":"Structural and sequence-based classification of glycoside hydrolases","volume":"7","author":"Henrissat B","year":"1997","unstructured":"B Henrissat, GJ Davies, Structural and sequence-based classification of glycoside hydrolases. Curr Opin Struct Biol 7, 637\u2013644 (1997).","journal-title":"Curr Opin Struct Biol"},{"key":"e_1_3_4_14_2","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.jmb.2003.10.026","article-title":"Crystal structure of \u03b2-D-xylosidase from Thermoanaerobacterium saccharolyticum, a family 39 glycoside hydrolase","volume":"335","author":"Yang JK","year":"2004","unstructured":"JK Yang, et al., Crystal structure of \u03b2-D-xylosidase from Thermoanaerobacterium saccharolyticum, a family 39 glycoside hydrolase. J Mol Biol 335, 155\u2013165 (2004).","journal-title":"J Mol Biol"},{"key":"e_1_3_4_15_2","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1016\/j.jmb.2005.09.003","article-title":"Enzyme-substrate complex structures of a GH39 \u03b2-xylosidase from Geobacillus stearothermophilus","volume":"353","author":"Czjzek M","year":"2005","unstructured":"M Czjzek, et al., Enzyme-substrate complex structures of a GH39 \u03b2-xylosidase from Geobacillus stearothermophilus. J Mol Biol 353, 838\u2013846 (2005).","journal-title":"J Mol Biol"},{"key":"e_1_3_4_16_2","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.ymgme.2004.12.006","article-title":"A homology model for human \u03b1-l-iduronidase: Insights into human disease","volume":"85","author":"Rempel BP","year":"2005","unstructured":"BP Rempel, LA Clarke, SG Withers, A homology model for human \u03b1-l-iduronidase: Insights into human disease. Mol Genet Metab 85, 28\u201337 (2005).","journal-title":"Mol Genet Metab"},{"key":"e_1_3_4_17_2","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/0009-8981(79)90121-9","article-title":"A fluorometric assay using 4-methylumbelliferyl \u03b1-L-iduronide for the estimation of \u03b1-L-iduronidase activity and the detection of Hurler and Scheie syndromes","volume":"92","author":"Hopwood JJ","year":"1979","unstructured":"JJ Hopwood, V Muller, A Smithson, N Baggett, A fluorometric assay using 4-methylumbelliferyl \u03b1-L-iduronide for the estimation of \u03b1-L-iduronidase activity and the detection of Hurler and Scheie syndromes. Clin Chim Acta 92, 257\u2013265 (1979).","journal-title":"Clin Chim Acta"},{"key":"e_1_3_4_18_2","doi-asserted-by":"crossref","first-page":"3278","DOI":"10.1016\/j.bmc.2009.03.050","article-title":"Effects of detergents on the West Nile virus protease activity","volume":"17","author":"Ezgimen MD","year":"2009","unstructured":"MD Ezgimen, NH Mueller, T Teramoto, R Padmanabhan, Effects of detergents on the West Nile virus protease activity. Bioorg Med Chem 17, 3278\u20133282 (2009).","journal-title":"Bioorg Med Chem"},{"key":"e_1_3_4_19_2","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.1107\/S1744309112040432","article-title":"Crystallization, X-ray diffraction analysis and SIRAS phasing of human \u03b1-L-iduronidase","volume":"68","author":"Maita N","year":"2012","unstructured":"N Maita, H Taniguchi, H Sakuraba, Crystallization, X-ray diffraction analysis and SIRAS phasing of human \u03b1-L-iduronidase. Acta Crystallogr Sect F Struct Biol Cryst Commun 68, 1363\u20131366 (2012).","journal-title":"Acta Crystallogr Sect F Struct Biol Cryst Commun"},{"key":"e_1_3_4_20_2","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1111\/j.1432-1033.1985.tb09158.x","article-title":"Human \u03b1-L-iduronidase. 1. Purification, monoclonal antibody production, native and subunit molecular mass","volume":"152","author":"Clements PR","year":"1985","unstructured":"PR Clements, DA Brooks, GT Saccone, JJ Hopwood, Human \u03b1-L-iduronidase. 1. Purification, monoclonal antibody production, native and subunit molecular mass. Eur J Biochem 152, 21\u201328 (1985).","journal-title":"Eur J Biochem"},{"key":"e_1_3_4_21_2","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/j.jsb.2010.11.024","article-title":"Crystal structures of Trichoderma reesei \u03b2-galactosidase reveal conformational changes in the active site","volume":"174","author":"Maksimainen M","year":"2011","unstructured":"M Maksimainen, et al., Crystal structures of Trichoderma reesei \u03b2-galactosidase reveal conformational changes in the active site. J Struct Biol 174, 156\u2013163 (2011).","journal-title":"J Struct Biol"},{"key":"e_1_3_4_22_2","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1111\/j.1467-7652.2005.00166.x","article-title":"Synthesis of enzymatically active human \u03b1-L-iduronidase in Arabidopsis cgl (complex glycan-deficient) seeds","volume":"4","author":"Downing WL","year":"2006","unstructured":"WL Downing, et al., Synthesis of enzymatically active human \u03b1-L-iduronidase in Arabidopsis cgl (complex glycan-deficient) seeds. Plant Biotechnol J 4, 169\u2013181 (2006).","journal-title":"Plant Biotechnol J"},{"key":"e_1_3_4_23_2","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.jprot.2012.09.022","article-title":"Characterization of \u03b1-L-Iduronidase (Aldurazyme) and its complexes","volume":"80","author":"Jung G","year":"2013","unstructured":"G Jung, M Pabst, L Neumann, A Berger, G Lubec, Characterization of \u03b1-L-Iduronidase (Aldurazyme) and its complexes. J Proteomics 80, 26\u201333 (2013).","journal-title":"J Proteomics"},{"key":"e_1_3_4_24_2","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1016\/S0969-2126(01)00220-9","article-title":"Structures and mechanisms of glycosyl hydrolases","volume":"3","author":"Davies G","year":"1995","unstructured":"G Davies, B Henrissat, Structures and mechanisms of glycosyl hydrolases. Structure 3, 853\u2013859 (1995).","journal-title":"Structure"},{"key":"e_1_3_4_25_2","doi-asserted-by":"crossref","first-page":"8054","DOI":"10.1021\/bi034293v","article-title":"Family 39 \u03b1-l-iduronidases and \u03b2-D-xylosidases react through similar glycosyl-enzyme intermediates: Identification of the human iduronidase nucleophile","volume":"42","author":"Nieman CE","year":"2003","unstructured":"CE Nieman, et al., Family 39 \u03b1-l-iduronidases and \u03b2-D-xylosidases react through similar glycosyl-enzyme intermediates: Identification of the human iduronidase nucleophile. Biochemistry 42, 8054\u20138065 (2003).","journal-title":"Biochemistry"},{"key":"e_1_3_4_26_2","doi-asserted-by":"crossref","first-page":"E2189","DOI":"10.1002\/humu.21479","article-title":"IDUA mutational profiling of a cohort of 102 European patients with mucopolysaccharidosis type I: Identification and characterization of 35 novel \u03b1-L-iduronidase (IDUA) alleles","volume":"32","author":"Bertola F","year":"2011","unstructured":"F Bertola, et al., IDUA mutational profiling of a cohort of 102 European patients with mucopolysaccharidosis type I: Identification and characterization of 35 novel \u03b1-L-iduronidase (IDUA) alleles. Hum Mutat 32, E2189\u2013E2210 (2011).","journal-title":"Hum Mutat"},{"key":"e_1_3_4_27_2","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1146\/annurev.bi.54.070185.003215","article-title":"Assembly of asparagine-linked oligosaccharides","volume":"54","author":"Kornfeld R","year":"1985","unstructured":"R Kornfeld, S Kornfeld, Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem 54, 631\u2013664 (1985).","journal-title":"Annu Rev Biochem"},{"key":"e_1_3_4_28_2","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.tips.2011.12.004","article-title":"A 3D view of autotaxin","volume":"33","author":"Nishimasu H","year":"2012","unstructured":"H Nishimasu, R Ishitani, J Aoki, O Nureki, A 3D view of autotaxin. Trends Pharmacol Sci 33, 138\u2013145 (2012).","journal-title":"Trends Pharmacol Sci"},{"key":"e_1_3_4_29_2","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1016\/S0022-2836(03)00172-4","article-title":"The structure of bovine lysosomal \u03b1-mannosidase suggests a novel mechanism for low-pH activation","volume":"327","author":"Heikinheimo P","year":"2003","unstructured":"P Heikinheimo, et al., The structure of bovine lysosomal \u03b1-mannosidase suggests a novel mechanism for low-pH activation. J Mol Biol 327, 631\u2013644 (2003).","journal-title":"J Mol Biol"},{"key":"e_1_3_4_30_2","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1107\/S0907444902018036","article-title":"Automated main-chain model building by template matching and iterative fragment extension","volume":"59","author":"Terwilliger TC","year":"2003","unstructured":"TC Terwilliger, Automated main-chain model building by template matching and iterative fragment extension. Acta Crystallogr D Biol Crystallogr 59, 38\u201344 (2003).","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"e_1_3_4_31_2","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1107\/S0907444910007493","article-title":"Features and development of Coot","volume":"66","author":"Emsley P","year":"2010","unstructured":"P Emsley, B Lohkamp, WG Scott, K Cowtan, Features and development of Coot. Acta Crystallogr D Biol Crystallogr 66, 486\u2013501 (2010).","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"e_1_3_4_32_2","doi-asserted-by":"crossref","first-page":"2728","DOI":"10.1038\/nprot.2007.406","article-title":"Version 1.2 of the Crystallography and NMR system","volume":"2","author":"Br\u00fcnger AT","year":"2007","unstructured":"AT Br\u00fcnger, Version 1.2 of the Crystallography and NMR system. Nat Protoc 2, 2728\u20132733 (2007).","journal-title":"Nat Protoc"},{"key":"e_1_3_4_33_2","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1107\/S0907444911001314","article-title":"REFMAC5 for the refinement of macromolecular crystal structures","volume":"67","author":"Murshudov GN","year":"2011","unstructured":"GN Murshudov, et al., REFMAC5 for the refinement of macromolecular crystal structures. Acta Crystallogr D Biol Crystallogr 67, 355\u2013367 (2011).","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"e_1_3_4_34_2","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1093\/bioinformatics\/btm404","article-title":"Clustal W and Clustal X version 2.0","volume":"23","author":"Larkin MA","year":"2007","unstructured":"MA Larkin, et al., Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947\u20132948 (2007).","journal-title":"Bioinformatics"},{"key":"e_1_3_4_35_2","doi-asserted-by":"crossref","first-page":"3320","DOI":"10.1093\/nar\/gkg556","article-title":"ESPript\/ENDscript: Extracting and rendering sequence and 3D information from atomic structures of proteins","volume":"31","author":"Gouet P","year":"2003","unstructured":"P Gouet, X Robert, E Courcelle, ESPript\/ENDscript: Extracting and rendering sequence and 3D information from atomic structures of proteins. Nucleic Acids Res 31, 3320\u20133323 (2003).","journal-title":"Nucleic Acids Res"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.1306939110","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,6,7]],"date-time":"2022-06-07T06:21:04Z","timestamp":1654582864000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.1306939110"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,8,19]]},"references-count":35,"journal-issue":{"issue":"36","published-print":{"date-parts":[[2013,9,3]]}},"alternative-id":["10.1073\/pnas.1306939110"],"URL":"https:\/\/doi.org\/10.1073\/pnas.1306939110","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,8,19]]},"assertion":[{"value":"2013-08-19","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}