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. 1993 Jun;91(6):2393–2398. doi: 10.1172/JCI116472

Protective protein gene mutations in galactosialidosis.

M Shimmoto 1, Y Fukuhara 1, K Itoh 1, A Oshima 1, H Sakuraba 1, Y Suzuki 1
PMCID: PMC443297  PMID: 8514852

Abstract

Four different protective protein cDNA mutations, 146A-->G (Q49R), 193T-->C (W65R), 268-269TC-->CT (S90L), and 1184A-->G (Y395C), were identified in six Japanese galactosialidosis patients with various phenotypic manifestations, and another mutation, 746T-->A (Y249N), in a patient of French-German origin with an atypical clinical course. Y395C was a common mutation in four Japanese patients in infancy and childhood; two juvenile patients were compound heterozygotes of Y395C and another common mutation, SpDEx7, and the other two infants were compound heterozygotes of Y395C and mutant alleles other than SpDEx7. We confirmed these mutations in genomic DNA by direct-sequence analysis or restriction-site analysis. The mutant cDNA clones, transiently expressed in a transformed galactosialidosis cell line, did not restore the secondarily deficient beta-galactosidase or alpha-neuraminidase activity except for the Y249N mutation that expressed some carboxypeptidase activity and restored the two lysosomal enzyme activities. Pulse-chase analysis detected a small amount of the mature form, as well as the precursor, in the cells transfected with the Y249N cDNA. Only precursor proteins were detected, mature proteins not appearing for the other mutant cDNAs.

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Selected References

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  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Galjart N. J., Gillemans N., Harris A., van der Horst G. T., Verheijen F. W., Galjaard H., d'Azzo A. Expression of cDNA encoding the human "protective protein" associated with lysosomal beta-galactosidase and neuraminidase: homology to yeast proteases. Cell. 1988 Sep 9;54(6):755–764. doi: 10.1016/s0092-8674(88)90999-3. [DOI] [PubMed] [Google Scholar]
  3. Itoh K., Takiyama N., Nagao Y., Oshima A., Sakuraba H., Potier M., Suzuki Y. Acid carboxypeptidase deficiency in galactosialidosis. Jinrui Idengaku Zasshi. 1991 Jun;36(2):171–177. doi: 10.1007/BF01876581. [DOI] [PubMed] [Google Scholar]
  4. Kase R., Itoh K., Takiyama N., Oshima A., Sakuraba H., Suzuki Y. Galactosialidosis: simultaneous deficiency of esterase, carboxy-terminal deamidase and acid carboxypeptidase activities. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1175–1179. doi: 10.1016/0006-291x(90)91572-a. [DOI] [PubMed] [Google Scholar]
  5. Miyazaki J., Takaki S., Araki K., Tashiro F., Tominaga A., Takatsu K., Yamamura K. Expression vector system based on the chicken beta-actin promoter directs efficient production of interleukin-5. Gene. 1989 Jul 15;79(2):269–277. doi: 10.1016/0378-1119(89)90209-6. [DOI] [PubMed] [Google Scholar]
  6. Nanba E., Tsuji A., Omura K., Suzuki Y. GM1-gangliosidosis: abnormalities in biosynthesis and early processing of beta-galactosidase in fibroblasts. Biochem Biophys Res Commun. 1988 Apr 29;152(2):794–800. doi: 10.1016/s0006-291x(88)80108-6. [DOI] [PubMed] [Google Scholar]
  7. Nanba E., Tsuji A., Omura K., Suzuki Y. Galactosialidosis: a direct evidence that a 46-kilodalton protein restores deficient enzyme activities in fibroblasts. Biochem Biophys Res Commun. 1987 Apr 14;144(1):138–142. doi: 10.1016/s0006-291x(87)80486-2. [DOI] [PubMed] [Google Scholar]
  8. Sakuraba H., Aoyagi T., Suzuki Y. Galactosialidosis (beta-galactosidase-neuraminidase deficiency): a possible role of serine-thiol proteases in the degradation of beta-galactosidase molecules. Clin Chim Acta. 1982 Nov 10;125(3):275–282. doi: 10.1016/0009-8981(82)90257-1. [DOI] [PubMed] [Google Scholar]
  9. Strisciuglio P., Sly W. S., Dodson W. E., McAlister W. H., Martin T. C. Combined deficiency of beta-galactosidase and neuraminidase: natural history of the disease in the first 18 years of an American patient with late infantile onset form. Am J Med Genet. 1990 Dec;37(4):573–577. doi: 10.1002/ajmg.1320370431. [DOI] [PubMed] [Google Scholar]
  10. Tranchemontagne J., Michaud L., Potier M. Deficient lysosomal carboxypeptidase activity in galactosialidosis. Biochem Biophys Res Commun. 1990 Apr 16;168(1):22–29. doi: 10.1016/0006-291x(90)91669-j. [DOI] [PubMed] [Google Scholar]
  11. Van Doren K., Gluzman Y. Efficient transformation of human fibroblasts by adenovirus-simian virus 40 recombinants. Mol Cell Biol. 1984 Aug;4(8):1653–1656. doi: 10.1128/mcb.4.8.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Verheijen F. W., Palmeri S., Hoogeveen A. T., Galjaard H. Human placental neuraminidase. Activation, stabilization and association with beta-galactosidase and its protective protein. Eur J Biochem. 1985 Jun 3;149(2):315–321. doi: 10.1111/j.1432-1033.1985.tb08928.x. [DOI] [PubMed] [Google Scholar]
  13. Wenger D. A., Tarby T. J., Wharton C. Macular cherry-red spots and myoclonus with dementia: coexistent neuraminidase and beta-galactosidase deficiencies. Biochem Biophys Res Commun. 1978 May 30;82(2):589–595. doi: 10.1016/0006-291x(78)90915-4. [DOI] [PubMed] [Google Scholar]
  14. Zhou X. Y., Galjart N. J., Willemsen R., Gillemans N., Galjaard H., d'Azzo A. A mutation in a mild form of galactosialidosis impairs dimerization of the protective protein and renders it unstable. EMBO J. 1991 Dec;10(13):4041–4048. doi: 10.1002/j.1460-2075.1991.tb04980.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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