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. 1997 May 15;99(10):2530–2537. doi: 10.1172/JCI119437

Identification and expression of acid beta-glucosidase mutations causing severe type 1 and neurologic type 2 Gaucher disease in non-Jewish patients.

M E Grace 1, R J Desnick 1, G M Pastores 1
PMCID: PMC508094  PMID: 9153297

Abstract

Gaucher disease, the most prevalent lysosomal storage disease, occurs in three subtypes, all resulting from mutations in the acid beta-glucosidase gene. Molecular studies in five severely affected type 1 and two type 2 Gaucher disease patients of non-Jewish descent identified six new mutations: K74X, W179X, G195E, S271N, V352L, and a two-base deletion in exon 10 (1450del2). Two additional mutations identified in these patients (R48W and G202R) have been reported previously, but were not expressed or characterized. Heterologous expression in Sf 9 cells using the baculovirus system revealed that the missense mutations, R48W and V352L, had 14 and 7%, respectively, of the specific activity based on cross-reacting immunologic material expressed by the normal allele. In contrast, the G195E, G202R, and S271N mutant alleles were more severely compromised with only 1-2% of the normal expressed specific activity based on cross-reacting immunologic material. Structural distortion at the active site was probed by comparing the interaction of the mutant enzymes with active site-directed inhibitors (castanospermine, conduritol B epoxide and deoxynojirimycin). R48W, G202R, and S271N were normally inhibited, whereas the V352L and G195E mutant enzymes had significantly decreased binding affinity. These mutations further expand the genetic heterogeneity in the lesions causing Gaucher disease types 1 and 2, and further delineate genotype/phenotype correlations and functional domains within the acid beta-glucosidase gene.

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Selected References

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  1. Barker D., Schafer M., White R. Restriction sites containing CpG show a higher frequency of polymorphism in human DNA. Cell. 1984 Jan;36(1):131–138. doi: 10.1016/0092-8674(84)90081-3. [DOI] [PubMed] [Google Scholar]
  2. Barneveld R. A., Keijzer W., Tegelaers F. P., Ginns E. I., Geurts van Kessel A., Brady R. O., Barranger J. A., Tager J. M., Galjaard H., Westerveld A. Assignment of the gene coding for human beta-glucocerebrosidase to the region q21-q31 of chromosome 1 using monoclonal antibodies. Hum Genet. 1983;64(3):227–231. doi: 10.1007/BF00279398. [DOI] [PubMed] [Google Scholar]
  3. Berg-Fussman A., Grace M. E., Ioannou Y., Grabowski G. A. Human acid beta-glucosidase. N-glycosylation site occupancy and the effect of glycosylation on enzymatic activity. J Biol Chem. 1993 Jul 15;268(20):14861–14866. [PubMed] [Google Scholar]
  4. Beutler E., Demina A., Gelbart T. Glucocerebrosidase mutations in Gaucher disease. Mol Med. 1994 Nov;1(1):82–92. [PMC free article] [PubMed] [Google Scholar]
  5. Beutler E. Gaucher disease as a paradigm of current issues regarding single gene mutations of humans. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5384–5390. doi: 10.1073/pnas.90.12.5384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beutler E. Gaucher disease: new molecular approaches to diagnosis and treatment. Science. 1992 May 8;256(5058):794–799. doi: 10.1126/science.1589760. [DOI] [PubMed] [Google Scholar]
  7. Beutler E., Gelbart T., Demina A., Zimran A., LeCoutre P. Five new Gaucher disease mutations. Blood Cells Mol Dis. 1995;21(1):20–24. doi: 10.1006/bcmd.1995.0004. [DOI] [PubMed] [Google Scholar]
  8. Beutler E., Gelbart T. Gaucher disease associated with a unique KpnI restriction site: identification of the amino-acid substitution. Ann Hum Genet. 1990 May;54(Pt 2):149–153. doi: 10.1111/j.1469-1809.1990.tb00371.x. [DOI] [PubMed] [Google Scholar]
  9. Beutler E., Gelbart T. Hematologically important mutations: Gaucher disease. Blood Cells Mol Dis. 1997;23(1):2–7. doi: 10.1006/bcmd.1997.0114. [DOI] [PubMed] [Google Scholar]
  10. Beutler E., West C., Gelbart T. Polymorphisms in the human glucocerebrosidase gene. Genomics. 1992 Apr;12(4):795–800. doi: 10.1016/0888-7543(92)90311-f. [DOI] [PubMed] [Google Scholar]
  11. Bordo D., Argos P. Suggestions for "safe" residue substitutions in site-directed mutagenesis. J Mol Biol. 1991 Feb 20;217(4):721–729. doi: 10.1016/0022-2836(91)90528-e. [DOI] [PubMed] [Google Scholar]
  12. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  13. Devine E. A., Smith M., Arredondo-Vega F. X., Shafit-Zagardo B., Desnick R. J. Chromosomal localization of the gene for Gaucher disease. Prog Clin Biol Res. 1982;95:511–534. [PubMed] [Google Scholar]
  14. Dinur T., Grabowski G. A., Desnick R. J., Gatt S. Synthesis of a fluorescent derivative of glucosyl ceramide for the sensitive determination of glucocerebrosidase activity. Anal Biochem. 1984 Jan;136(1):223–234. doi: 10.1016/0003-2697(84)90329-4. [DOI] [PubMed] [Google Scholar]
  15. Ginns E. I., Choudary P. V., Tsuji S., Martin B., Stubblefield B., Sawyer J., Hozier J., Barranger J. A. Gene mapping and leader polypeptide sequence of human glucocerebrosidase: implications for Gaucher disease. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7101–7105. doi: 10.1073/pnas.82.20.7101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grace M. E., Berg A., He G. S., Goldberg L., Horowitz M., Grabowski G. A. Gaucher disease: heterologous expression of two alleles associated with neuronopathic phenotypes. Am J Hum Genet. 1991 Sep;49(3):646–655. [PMC free article] [PubMed] [Google Scholar]
  17. Grace M. E., Graves P. N., Smith F. I., Grabowski G. A. Analyses of catalytic activity and inhibitor binding of human acid beta-glucosidase by site-directed mutagenesis. Identification of residues critical to catalysis and evidence for causality of two Ashkenazi Jewish Gaucher disease type 1 mutations. J Biol Chem. 1990 Apr 25;265(12):6827–6835. [PubMed] [Google Scholar]
  18. Grace M. E., Newman K. M., Scheinker V., Berg-Fussman A., Grabowski G. A. Analysis of human acid beta-glucosidase by site-directed mutagenesis and heterologous expression. J Biol Chem. 1994 Jan 21;269(3):2283–2291. [PubMed] [Google Scholar]
  19. Grantham R. Amino acid difference formula to help explain protein evolution. Science. 1974 Sep 6;185(4154):862–864. doi: 10.1126/science.185.4154.862. [DOI] [PubMed] [Google Scholar]
  20. He G. S., Grace M. E., Grabowski G. A. Gaucher disease: four rare alleles encoding F213I, P289L, T323I, and R463C in type 1 variants. Hum Mutat. 1992;1(5):423–427. doi: 10.1002/humu.1380010513. [DOI] [PubMed] [Google Scholar]
  21. Henrissat B., Callebaut I., Fabrega S., Lehn P., Mornon J. P., Davies G. Conserved catalytic machinery and the prediction of a common fold for several families of glycosyl hydrolases. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7090–7094. doi: 10.1073/pnas.92.15.7090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ho M. W., O'Brien J. S. Gaucher's disease: deficiency of 'acid' -glucosidase and reconstitution of enzyme activity in vitro. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2810–2813. doi: 10.1073/pnas.68.11.2810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Holliday R., Grigg G. W. DNA methylation and mutation. Mutat Res. 1993 Jan;285(1):61–67. doi: 10.1016/0027-5107(93)90052-h. [DOI] [PubMed] [Google Scholar]
  24. Horowitz M., Wilder S., Horowitz Z., Reiner O., Gelbart T., Beutler E. The human glucocerebrosidase gene and pseudogene: structure and evolution. Genomics. 1989 Jan;4(1):87–96. doi: 10.1016/0888-7543(89)90319-4. [DOI] [PubMed] [Google Scholar]
  25. Horowitz M., Zimran A. Mutations causing Gaucher disease. Hum Mutat. 1994;3(1):1–11. doi: 10.1002/humu.1380030102. [DOI] [PubMed] [Google Scholar]
  26. Hutchison C. A., 3rd, Swanstrom R., Loeb D. D. Complete mutagenesis of protein coding domains. Methods Enzymol. 1991;202:356–390. doi: 10.1016/0076-6879(91)02019-6. [DOI] [PubMed] [Google Scholar]
  27. Kawame H., Eto Y. A new glucocerebrosidase-gene missense mutation responsible for neuronopathic Gaucher disease in Japanese patients. Am J Hum Genet. 1991 Dec;49(6):1378–1380. [PMC free article] [PubMed] [Google Scholar]
  28. Kim J. W., Liou B. B., Lai M. Y., Ponce E., Grabowski G. A. Gaucher disease: identification of three new mutations in the Korean and Chinese (Taiwanese) populations. Hum Mutat. 1996;7(3):214–218. doi: 10.1002/(SICI)1098-1004(1996)7:3<214::AID-HUMU5>3.0.CO;2-A. [DOI] [PubMed] [Google Scholar]
  29. Miao S., McCarter J. D., Grace M. E., Grabowski G. A., Aebersold R., Withers S. G. Identification of Glu340 as the active-site nucleophile in human glucocerebrosidase by use of electrospray tandem mass spectrometry. J Biol Chem. 1994 Apr 15;269(15):10975–10978. [PubMed] [Google Scholar]
  30. Roitsch T., Lehle L. Structural requirements for protein N-glycosylation. Influence of acceptor peptides on cotranslational glycosylation of yeast invertase and site-directed mutagenesis around a sequon sequence. Eur J Biochem. 1989 May 1;181(2):525–529. doi: 10.1111/j.1432-1033.1989.tb14755.x. [DOI] [PubMed] [Google Scholar]
  31. Sanger F. Determination of nucleotide sequences in DNA. Science. 1981 Dec 11;214(4526):1205–1210. doi: 10.1126/science.7302589. [DOI] [PubMed] [Google Scholar]
  32. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  33. Sibille A., Eng C. M., Kim S. J., Pastores G., Grabowski G. A. Phenotype/genotype correlations in Gaucher disease type I: clinical and therapeutic implications. Am J Hum Genet. 1993 Jun;52(6):1094–1101. [PMC free article] [PubMed] [Google Scholar]
  34. Sorge J., Gross E., West C., Beutler E. High level transcription of the glucocerebrosidase pseudogene in normal subjects and patients with Gaucher disease. J Clin Invest. 1990 Oct;86(4):1137–1141. doi: 10.1172/JCI114818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sorge J., West C., Westwood B., Beutler E. Molecular cloning and nucleotide sequence of human glucocerebrosidase cDNA. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7289–7293. doi: 10.1073/pnas.82.21.7289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sorge J., West C., Westwood B., Beutler E. Molecular cloning and nucleotide sequence of human glucocerebrosidase cDNA. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7289–7293. doi: 10.1073/pnas.82.21.7289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Taylor J. W., Ott J., Eckstein F. The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Res. 1985 Dec 20;13(24):8765–8785. doi: 10.1093/nar/13.24.8765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Taylor J. W., Schmidt W., Cosstick R., Okruszek A., Eckstein F. The use of phosphorothioate-modified DNA in restriction enzyme reactions to prepare nicked DNA. Nucleic Acids Res. 1985 Dec 20;13(24):8749–8764. doi: 10.1093/nar/13.24.8749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Theophilus B., Latham T., Grabowski G. A., Smith F. I. Gaucher disease: molecular heterogeneity and phenotype-genotype correlations. Am J Hum Genet. 1989 Aug;45(2):212–225. [PMC free article] [PubMed] [Google Scholar]
  40. Tsuji S., Choudary P. V., Martin B. M., Stubblefield B. K., Mayor J. A., Barranger J. A., Ginns E. I. A mutation in the human glucocerebrosidase gene in neuronopathic Gaucher's disease. N Engl J Med. 1987 Mar 5;316(10):570–575. doi: 10.1056/NEJM198703053161002. [DOI] [PubMed] [Google Scholar]
  41. Tsuji S., Choudary P. V., Martin B. M., Winfield S., Barranger J. A., Ginns E. I. Nucleotide sequence of cDNA containing the complete coding sequence for human lysosomal glucocerebrosidase. J Biol Chem. 1986 Jan 5;261(1):50–53. [PubMed] [Google Scholar]
  42. Wang Q., Trimbur D., Graham R., Warren R. A., Withers S. G. Identification of the acid/base catalyst in Agrobacterium faecalis beta-glucosidase by kinetic analysis of mutants. Biochemistry. 1995 Nov 7;34(44):14554–14562. doi: 10.1021/bi00044a034. [DOI] [PubMed] [Google Scholar]
  43. Zimran A., Sorge J., Gross E., Kubitz M., West C., Beutler E. Prediction of severity of Gaucher's disease by identification of mutations at DNA level. Lancet. 1989 Aug 12;2(8659):349–352. doi: 10.1016/s0140-6736(89)90536-9. [DOI] [PubMed] [Google Scholar]

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