Abstract
We produced six transgenic mouse lines expressing human α-galactosidase (α-Gal) in order to evaluate its posttranslational modification. Among them, serum α-Gal activity increased 3000-fold in two transgenic mouse lines (TgN2 and TgN51), as compared to that in non-transgenic lines. The heart and liver of the TgN2 mouse expressed a high amount of transcript as well as high α-Gal activity. Its gene products in the heart and kidney were sensitive to endoglycosidase H digestion, but those in the spleen and liver were largely resistant. Glycopeptidase F treatment confirmed an identical molecular mass for the peptide moiety of the enzyme. We concluded that heterogeneous molecular mass of the gene products was caused by different degrees of posttranslational glycosylation in murine tissues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Desnick RJ, Ioannou YA, Eng CM (1995) α-Galactosidase A deficiency: Fabry disease In The Metabolic and Molecular Bases of Inherited Disease, 7th edition (Scriver CR, Beaudet AI, Sly WS, Valle D, eds) pp 2741–84. New York: McGraw-Hill.
Fukuhara Y, Sakuraba H, Oshima A, Shimmoto M, Nagao Y, Nadaoka Y, Suzuki T, Suzuki Y (1990) Partial deletion of human α-galactosidase A gene in Fabry disease: Direct repeat sequences as a possible cause of slipped mispairing. Biochem Biophys Res Commun 170: 296–300.
Kornreich R, Bishop DF, Desnick RJ (1990) α-Galactosidase A gene rearrangements causing Fabry disease: Identification of short direct repeats at breakpoints in an Alu-rich gene. J Biol Chem 265: 9319–26.
Ishii S, Sakuraba H, Shimmoto M, Minamikawa-Tachino R, Suzuki T, Suzuki Y (1991) Fabry disease: Detection of 13–bp deletion in α-galactosidase A gene and its application to gene diagnosis of heterozygotes. Ann Neurol 29: 560–4.
Davies JP, Winchester BG, Malcolm S (1993) Mutation analysis in patients with the typical form of Anderson-Fabry disease. Hum Molec Genet 2: 1051–3.
Eng CM, Resnick-Silverman LA, Niehaus DJ, Astrin KH, Desnick RJ (1993) Nature and frequency of mutations in the α-galactosidase A gene that cause Fabry disease. Am J Hum Genet 53: 1186–97.
Sakuraba H, Eng CM, Desnick RJ, Bishop DF (1992) Invariant exon skipping in the human α-galactosidase A pre-mRNA: A g+1 to t substitution in 5′-splice site causing Fabry disease. Genomics 12: 643–50.
Okumiya T, Ishii S, Kase R, Kamei S, Sakuraba H, Suzuki Y (1995) α-galactosidase gene mutations in Fabry disease: heterogeneous expressions of mutant enzyme proteins. Hum Genet 95: 557–61.
Sakuraba H, Oshima A, Fukuhara Y, Shimmoto M, Nagao Y, Bishop DF, Desnick RJ, Suzuki Y (1990) Identification of point mutations in the α-galactosidase A gene in classical and atypical hemizygotes with Fabry disease. Am J Hum Genet 47: 784–9.
Koide T, Ishiura M, Iwai K, Inoue M, Kaneda Y, Uchida T (1990) A case of Fabry's disease in a patient with no α-galactosidase A activity caused by a single amino acid substitution of Pro-40 by Ser. FEBS Lett 259: 353–6.
Ishii S, Sakuraba H, Suzuki Y (1992) Point mutations in the upstream region of α-galactosidase A gene exon 6 in atypical variant of Fabry disease. Hum Genet 89: 29–32.
Nakao S, Takenaka T, Maeda M, Kodama C, Tanaka A, Tahara M, Yoshida A, Kuriyama M, Hayashibe H, SakurabaH, Tanaka H (1995) An atypical variant of Fabry's disease in men with left ventricular hypertrophy. N Engl J Med 333: 288–93.
Mapes CA, Anderson RL, Sweeley CC, Desnick RJ, Krivit W (1970) Enzyme replacement in Fabry's disease, an inborn error of metabolism. Science 169: 987–9.
Brady RO, Tallman JF, Johnson WG, Gal AE, Leahy WR, Quirk JM, Dekaban AS (1973) Replacement therapy for inherited enzyme deficiency: Use of purified ceramidetrihexosidase in Fabry's disease. N Engl J Med 289: 9–14.
Desnick RJ, Dean KJ, Grabowski GA, Bishop DF, Sweeley CC (1979) Enzyme therapy XII: Enzyme therapy in Fabry's disease: Differencial enzyme and substrate clearance kinetics of plasma and splenic α-galactosidase isozymes. Proc Natl Acad Sci USA 76: 5326–30.
Ishii S, Kase R, Sakuraba H, Suzuki Y (1993) Characterization of a mutant α-galactosidase gene product for the late-onset cardiac form of Fabry disease. Biochem Biophys Res Commun 197: 1585–9.
Brinster RL, Chen HY, Trumbauer MD, Yagle MK, Palmiter RD (1985) Fators affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc Natl Acad Sci USA 82: 4438–42.
Aldridge J, Kunkel L, Bruns G, Tantravahi U, Lalande M, Brewster T, Moreau E, Wilson M, Bromley W, Roderick T, Latt SA (1984) A strategy to reveal high-frequency RFLPs along the human X chromosome. Am J Hum Genet 36: 546–64.
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–9.
Ishii S, Kase R, Sakuraba H, Fujita S, Sugimoto M, Tomita K, Semba T, Suzuki Y (1994) Human α-galactosidase gene expression: significance of two peptide regions encoded by exons 1–2 and 6. Biochim Biophys Acta 1204: 265–70.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–54.
Làemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–5.
Bishop DF, Desnick RJ (1981) Affinity purification of α-galactosidase A from human spleen, placenta, and plasma with elimination of pyrogen contamination. Properties of the purified splenic enzyme compared to other forms. J Biol Chem 256: 1307–16.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ishii, S., Kase, R., Sakuraba, H. et al. α-Galactosidase transgenic mouse: Heterogeneous gene expression and posttranslational glycosylation in tissues. Glycoconj J 15, 591–594 (1998). https://doi.org/10.1023/A:1006915926732
Issue Date:
DOI: https://doi.org/10.1023/A:1006915926732