{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T23:03:33Z","timestamp":1792710213203,"version":"4.5.0"},"reference-count":50,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2007,9,1]]},"abstract":"<jats:p>Fabry disease is a lysosomal storage disorder caused by the deficiency of \u03b1-Gal A (\u03b1-galactosidase A) activity. In order to understand the molecular mechanism underlying \u03b1-Gal A deficiency in Fabry disease patients with residual enzyme activity, enzymes with different missense mutations were purified from transfected COS-7 cells and the biochemical properties were characterized. The mutant enzymes detected in variant patients (A20P, E66Q, M72V, I91T, R112H, F113L, N215S, Q279E, M296I, M296V and R301Q), and those found mostly in mild classic patients (A97V, A156V, L166V and R356W) appeared to have normal Km and Vmax values. The degradation of all mutants (except E59K) was partially inhibited by treatment with kifunensine, a selective inhibitor of ER (endoplasmic reticulum) \u03b1-mannosidase I. Metabolic labelling and subcellular fractionation studies in COS-7 cells expressing the L166V and R301Q \u03b1-Gal A mutants indicated that the mutant protein was retained in the ER and degraded without processing. Addition of DGJ (1-deoxygalactonojirimycin) to the culture medium of COS-7 cells transfected with a large set of missense mutant \u03b1-Gal A cDNAs effectively increased both enzyme activity and protein yield. DGJ was capable of normalizing intracellular processing of mutant \u03b1-Gal A found in both classic (L166V) and variant (R301Q) Fabry disease patients. In addition, the residual enzyme activity in fibroblasts or lymphoblasts from both classic and variant hemizygous Fabry disease patients carrying a variety of missense mutations could be substantially increased by cultivation of the cells with DGJ. These results indicate that a large proportion of mutant enzymes in patients with residual enzyme activity are kinetically active. Excessive degradation in the ER could be responsible for the deficiency of enzyme activity in vivo, and the DGJ approach may be broadly applicable to Fabry disease patients with missense mutations.<\/jats:p>","DOI":"10.1042\/bj20070479","type":"journal-article","created":{"date-parts":[[2007,8,14]],"date-time":"2007-08-14T08:40:13Z","timestamp":1187080813000},"page":"285-295","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":134,"title":["Mutant \u03b1-galactosidase A enzymes identified in Fabry disease patients with residual enzyme activity: biochemical characterization and restoration of normal intracellular processing by 1-deoxygalactonojirimycin"],"prefix":"10.1042","volume":"406","author":[{"given":"Satoshi","family":"Ishii","sequence":"first","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, Fifth Avenue at 100th Street, New York, NY 10029, U.S.A."},{"name":"Department of Agricultural and Life Sciences, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, 080-8555, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui-Hwa","family":"Chang","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, Fifth Avenue at 100th Street, New York, NY 10029, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kunito","family":"Kawasaki","sequence":"additional","affiliation":[{"name":"Department of Agricultural and Life Sciences, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, 080-8555, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kayo","family":"Yasuda","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, Fifth Avenue at 100th Street, New York, NY 10029, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui-Li","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, Fifth Avenue at 100th Street, New York, NY 10029, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott\u00a0C.","family":"Garman","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Massachusetts, 710 North Pleasant Street, Amherst, MA 01003, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian-Qiang","family":"Fan","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, Fifth Avenue at 100th Street, New York, NY 10029, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2007,8,13]]},"reference":[{"key":"2021112213292044300_B1","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1056\/NEJM196705252762101","article-title":"Enzymatic defect in Fabry's disease: ceramidetrihexosidase deficiency","volume":"276","author":"Brady","year":"1967","journal-title":"N. 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Med."},{"key":"2021112213292044300_B32","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1093\/hmg\/3.10.1795","article-title":"Fabry disease: twenty-three mutations including sense and antisense CpG alterations and identification of a deletional hot-spot in the \u03b1-galactosidase A gene","volume":"3","author":"Eng","year":"1994","journal-title":"Hum. Mol. Genet."},{"key":"2021112213292044300_B33","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1007\/BF00223869","article-title":"\u03b1-Galactosidase gene mutations in Fabry disease: heterogeneous expressions of mutant enzyme proteins","volume":"95","author":"Okumiya","year":"1995","journal-title":"Hum. Genet."},{"key":"2021112213292044300_B34","doi-asserted-by":"crossref","first-page":"1390","DOI":"10.1172\/JCI114027","article-title":"Fabry disease: six gene rearrangements and an exonic point mutation in the \u03b1-galactosidase gene","volume":"83","author":"Bernstein","year":"1989","journal-title":"J. Clin. Invest."},{"key":"2021112213292044300_B35","first-page":"227","article-title":"Fabry disease: twenty-two novel mutations in the \u03b1-galactosidase A gene and genotype\/phenotype correlations in severely and mildly affected hemizygotes and heterozygotes","volume":"48","author":"Ashton-Prolla","year":"2000","journal-title":"J. Investig. Med."},{"key":"2021112213292044300_B36","first-page":"784","article-title":"Identification of point mutations in the \u03b1-galactosidase A gene in classical and atypical hemizygotes with Fabry disease","volume":"47","author":"Sakuraba","year":"1990","journal-title":"Am. J. Hum. Genet."},{"key":"2021112213292044300_B37","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1002\/humu.41","article-title":"Identification of a novel de novo mutation (G373D) in the \u03b1-galactosidase A gene (GLA) in a patient affected with Fabry disease","volume":"17","author":"Germain","year":"2001","journal-title":"Hum. Mutat."},{"key":"2021112213292044300_B38","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1093\/hmg\/3.3.503","article-title":"Six novel mutations in the \u03b1-galactosidase A gene in families with Fabry disease","volume":"3","author":"Ploos\u00a0van\u00a0Amstel","year":"1994","journal-title":"Hum. Mol. Genet."},{"key":"2021112213292044300_B39","doi-asserted-by":"crossref","first-page":"7195","DOI":"10.1016\/S0021-9258(19)44613-9","article-title":"Purification and properties of human \u03b1-galactosidases","volume":"247","author":"Beutler","year":"1972","journal-title":"J. Biol. Chem."},{"key":"2021112213292044300_B40","doi-asserted-by":"crossref","first-page":"2062","DOI":"10.1016\/S0021-9258(18)61618-7","article-title":"Synthesis and processing of \u03b1-galactosidase A in human fibroblasts. Evidence for different mutations in Fabry disease","volume":"262","author":"Lemansky","year":"1987","journal-title":"J. Biol. 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U.S.A."},{"key":"2021112213292044300_B44","doi-asserted-by":"crossref","first-page":"4082","DOI":"10.1111\/j.1742-4658.2006.05410.x","article-title":"Hydrophilic iminosugar active-site-specific chaperones increase residual glucocerebrosidase activity in fibroblasts from Gaucher patients","volume":"273","author":"Chang","year":"2006","journal-title":"FEBS J."},{"key":"2021112213292044300_B45","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1007\/BF00219337","article-title":"Quantitative correlation between the residual activity of \u03b2-hexosaminidase A and arylsulfatase A and the severity of the resulting lysosomal storage disease","volume":"88","author":"Leinekugel","year":"1992","journal-title":"Hum. 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