Skip to main content
Log in

Mutation of the GM2 activator protein in a feline model of GM2 gangliosidosis

  • Regular Paper
  • Published:
Acta Neuropathologica Aims and scope Submit manuscript

Abstract

The GM2 activator protein is required for successful degradation of GM2 ganglioside by the A isozyme of lysosomal β-N-acetylhexosaminidase (EC 3.2.1.52). Deficiency of the GM2 activator protein leads to a relentlessly progressive accumulation of GM2 ganglioside in neuronal lysosomes and subsequent fatal deterioration of central nervous system function. GM2 activator deficiency has been described in humans, dogs and mice. This manuscript reports the discovery and characterization of a feline model of GM2 activator deficiency that exhibits many disease traits typical of the disorder in other species. Cats deficient in the GM2 activator protein develop clinical signs at approximately 14 months of age, including motor incoordination and exaggerated startle response to sharp sounds. Affected cats exhibit central nervous system abnormalities such as swollen neurons, membranous cytoplasmic bodies, increased sialic acid content and elevated levels of GM2 ganglioside. As is typical of GM2 activator deficiency, hexosaminidase A activity in tissue homogenates appears normal when assayed with a commonly used synthetic substrate. When the GM2 activator cDNA was sequenced from normal and affected cats, a deletion of 4 base pairs was identified as the causative mutation, resulting in alteration of 21 amino acids at the C terminus of the GM2 activator protein.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Baker HJ, Lindsey JR, McKhann GM, Farrell DF (1971) Neuronal GM1 gangliosidosis in a Siamese cat with beta-galactosidase deficiency. Science 174:838–839

    PubMed  Google Scholar 

  2. Baker HJ, Reynolds GD, Walkley SU, Cox NR, Baker GH (1979) The gangliosidoses: comparative features and research applications. Vet Pathol 16:635–649

    PubMed  Google Scholar 

  3. Bellachioma G, Stirling JL, Orlacchio A, Beccari T (1993) Cloning and sequence analysis of a cDNA clone coding for the mouse GM2 activator protein. Biochem J 294:227–230

    PubMed  Google Scholar 

  4. Bertoni C, Appolloni MG, Stirling JL, Li SC, Li YT, Orlacchio A, Beccari T (1997) Structural organization and expression of the gene for the mouse GM2 activator protein. Mamm Genome 8:90–93

    Article  PubMed  Google Scholar 

  5. Bertoni C, Li YT, Li SC (1999) Catabolism of asialo-GM2 in man and mouse. Specificity of human/mouse chimeric GM2 activator proteins. J Biol Chem 274:28612–28618

    Article  PubMed  Google Scholar 

  6. Chen B, Rigat B, Curry C, Mahuran DJ (1999) Structure of the GM2A gene: identification of an exon 2 nonsense mutation and a naturally occurring transcript with an in-frame deletion of exon 2. Am J Hum Genet 65:77–87

    Article  PubMed  Google Scholar 

  7. Conzelmann E, Sandhoff K (1978) AB variant of infantile GM2 gangliosidosis: deficiency of a factor necessary for stimulation of hexosaminidase A-catalyzed degradation of ganglioside GM2 and glycolipid GA2. Proc Natl Acad Sci USA 75:3979–3983

    PubMed  Google Scholar 

  8. Cork LC, Munnell JF, Lorenz MD, Murphy JV, Baker HJ, Rattazzi MC (1977) GM2 ganglioside lysosomal storage disease in cats with beta-hexosaminidase deficiency. Science 196:1014–1017

    PubMed  Google Scholar 

  9. Cummings JF, Wood PA, Walkley SU, de Lahunta A, DeForest ME (1985) GM2 gangliosidosis in a Japanese spaniel. Acta Neuropathol 67:247–253

    Article  PubMed  Google Scholar 

  10. Frischmeyer PA, Dietz HC (1999) Nonsense-mediated mRNA decay in health and disease. Hum Mol Genet 8:1893–1900

    Article  PubMed  Google Scholar 

  11. Gravel RA, Kaback MM, Proia RL, Sandhoff K, Suzuki K, Suzuki K (2001) The GM2 gangliosidoses. In: Scriver CR, Sly WS, Childs B, Beaudet AL, Valle D, Kinzler KW, Vogelstein B (eds) The metabolic and molecular bases of inherited disease, 8th edn. McGraw-Hill, New York, pp 3827–3876

  12. Guidotti JE, Mignon A, Haase G, Caillaud C, McDonell N, Kahn A, Poenaru L (1999) Adenoviral gene therapy of the Tay-Sachs disease in hexosaminidase A-deficient knock-out mice. Hum Mol Genet 8:831–838

    Article  PubMed  Google Scholar 

  13. Ho TT, Maguire AM, Aguirre GD, Surace EM, Anand V, Zeng Y, Salvetti A, Hopwood JJ, Haskins ME, Bennett J (2002) Phenotypic rescue after adeno-associated virus-mediated delivery of 4-sulfatase to the retinal pigment epithelium of feline mucopolysaccharidosis VI. J Gene Med 4:613–621

    Article  PubMed  Google Scholar 

  14. Ishikawa Y, Li SC, Wood PA, Li YT (1987) Biochemical basis of type AB GM2 gangliosidosis in a Japanese spaniel. J Neurochem 48:860–864

    PubMed  Google Scholar 

  15. Jeyakumar M, Butters TD, Cortina-Borja M, Hunnam V, Proia RL, Perry VH, Dwek RA, Platt FM (1999) Delayed symptom onset and increased life expectancy in Sandhoff disease mice treated with N-butyldeoxynojirimycin. Proc Natl Acad Sci USA 96:6388–6393

    Article  PubMed  Google Scholar 

  16. Jeyakumar M, Norflus F, Tifft CJ, Cortina-Borja M, Butters TD, Proia RL, Perry VH, Dwek RA, Platt FM (2001) Enhanced survival in Sandhoff disease mice receiving a combination of substrate deprivation therapy and bone marrow transplantation. Blood 97:327–329

    Article  PubMed  Google Scholar 

  17. Jeyakumar M, Smith D, Eliott-Smith E, Cortina-Borja M, Reinkensmeier G, Butters TD, Lemm T, Sandhoff K, Perry VH, Dwek RA, Platt FM (2002) An inducible mouse model of late onset Tay-Sachs disease. Neurobiol Dis 10:201–210

    Article  PubMed  Google Scholar 

  18. Kundu SK, Suzuki A (1981) Simple micro-method for the isolation of gangliosides by reversed-phase chromatography. J Chromatogr B Biomed Sci Appl 224:249-256

    Article  Google Scholar 

  19. Liu Y, Hoffmann A, Grinberg A, Westphal H, McDonald MP, Miller KM, Crawley JN, Sandhoff K, Suzuki K, Proia RL (1997) Mouse model of GM2 activator deficiency manifests cerebellar pathology and motor impairment. Proc Natl Acad Sci USA 94:8138–8143

    Article  PubMed  Google Scholar 

  20. Mahuran DJ (1998) The GM2 activator protein, its roles as a co-factor in GM2 hydrolysis and as a general glycolipid transport protein. Biochim Biophys Acta 1393:1–18

    PubMed  Google Scholar 

  21. Martin DR, Cox NR, Hathcock TL, Niemeyer GP, Baker HJ (2002) Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow. Exp Hematol 30:879–886

    Article  PubMed  Google Scholar 

  22. Martin DR, Krum BK, Varadarajan GS, Hathcock TL, Smith BF, Baker HJ (2004) An inversion of 25 base pairs causes feline GM2 gangliosidosis variant 0. Exp Neurol 187:30–37

    Article  PubMed  Google Scholar 

  23. Miettinen T (1959) Use of butyl acetate in determination of sialic acid. Acta Chem Scand 13:856–858

    Google Scholar 

  24. Neuwelt EA, Johnson WG, Blank NK, Pagel MA, Maslen-McClure C, McClure MJ, Wu PM (1985) Characterization of a new model of GM2-gangliosidosis (Sandhoff’s disease) in Korat cats. J Clin Invest 76:482–490

    PubMed  Google Scholar 

  25. Peterson GL (1977) A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem 83:346–356

    Article  PubMed  Google Scholar 

  26. Platt FM, Neises GR, Reinkensmeier G, Townsend MJ, Perry VH, Proia RL, Winchester B, Dwek RA, Butters TD (1997) Prevention of lysosomal storage in Tay-Sachs mice treated with N-butyldeoxynojirimycin. Science 276:428–431

    Article  PubMed  Google Scholar 

  27. Rattazzi MC, Lanse SB, McCullough RA, Nester JA, Jacobs EA (1980) Towards enzyme replacement in GM2 gangliosidosis: organ disposition and induced central nervous system uptake of human beta-hexosaminidase in the cat. Birth Defects Orig Artic Ser 16:179–193

    Google Scholar 

  28. Reynolds GD, Baker HJ, Reynolds RH (1978) Enzyme replacement using liposome carriers in feline GM1 gangliosidosis fibroblasts. Nature 275:754–755

    Article  PubMed  Google Scholar 

  29. Sango K, Yamanaka S, Hoffmann A, Okuda Y, Grinberg A, Westphal H, McDonald MP, Crawley JN, Sandhoff K, Suzuki K (1995) Mouse models of Tay-Sachs and Sandhoff diseases differ in neurologic phenotype and ganglioside metabolism. Nat Genet 11:170–176

    Article  PubMed  Google Scholar 

  30. Smiljanic-Georgijev N, Rigat B, Xie B, Wang W, Mahuran DJ (1997) Characterization of the affinity of the G(M2) activator protein for glycolipids by a fluorescence dequenching assay. Biochim Biophys Acta 1339:192–202

    PubMed  Google Scholar 

  31. Svennerholm L (1957) Quantitative estimation of sialic acids II. A colorimetric resorcinol-hydrochloric acid method. Biochim Biophys Acta 24:604–611

    Article  PubMed  Google Scholar 

  32. Vite CH, Passini MA, Haskins ME, Wolfe JH (2003) Adeno-associated virus vector-mediated transduction in the cat brain. Gene Ther 10:1874–1881

    Article  PubMed  Google Scholar 

  33. Wada R, Tifft CJ, Proia RL (2000) Microglial activation precedes acute neurodegeneration in Sandhoff disease and is suppressed by bone marrow transplantation. Proc Natl Acad Sci USA 97:10954–10959

    Article  PubMed  Google Scholar 

  34. Walkley SU, Thrall MA, Dobrenis K, Huang M, March PA, Siegel DA, Wurzelmann S (1994) Bone marrow transplantation corrects the enzyme defect in neurons of the central nervous system in a lysosomal storage disease. Proc Natl Acad Sci USA 91:2970–2974

    PubMed  Google Scholar 

  35. Wendeler M, Hoernschemeyer J, Hoffmann D, Kolter T, Schwarzmann G, Sandhoff K (2004) Photoaffinity labelling of the human GM2-activator protein. Mechanistic insight into ganglioside GM2 degradation. Eur J Biochem 271:614–627

    Article  PubMed  Google Scholar 

  36. Wright CS, Li SC, Rastinejad F (2000) Crystal structure of human GM2-activator protein with a novel beta-cup topology. J Mol Biol 304:411–422

    Article  PubMed  Google Scholar 

  37. Wright CS, Zhao Q, Rastinejad F (2003) Structural analysis of lipid complexes of GM2-activator protein. J Mol Biol 331:951–964

    Article  PubMed  Google Scholar 

  38. Xie B, Kennedy JL, McInnes B, Auger D, Mahuran D (1992) Identification of a processed pseudogene related to the functional gene encoding the GM2 activator protein: localization of the pseudogene to human chromosome 3 and the functional gene to human chromosome 5. Genomics 14:796–798

    Article  PubMed  Google Scholar 

  39. Xie B, Rigat B, Smiljanic-Georgijev N, Deng H, Mahuran D (1998) Biochemical characterization of the Cys138Arg substitution associated with the AB variant form of GM2 gangliosidosis: evidence that Cys138 is required for the recognition of the GM2 activator/GM2 ganglioside complex by beta-hexosaminidase A. Biochemistry 37:814–821

    Article  PubMed  Google Scholar 

  40. Yuziuk JA, Bertoni C, Beccari T, Orlacchio A, Wu YY, Li SC, Li YT (1998) Specificity of mouse GM2 activator protein and beta-N-acetylhexosaminidases A and B. Similarities and differences with their human counterparts in the catabolism of GM2. J Biol Chem 273:66–72

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The authors acknowledge the assistance of Dr. R. French, pathologist at the Veterinary Diagnostic Laboratory, University of Illinois at Urbana for performing the necropsy and initial histological description of the proband. Electron microscopy was performed expertly by M. A. Toivio-Kinnucan, Department of Pathobiology, Auburn University College of Veterinary Medicine. This research was funded by the Scott-Ritchey Research Center, Auburn, AL. Accession nos.: GM2 activator cDNAs from normal and affected cats were sequenced and deposited in the GenBank database under accession numbers AY553653 (feline normal) and AY553654 (feline affected). Additional GM2 activator cDNAs referenced in this manuscript are L01439 (human) and L19526 (mouse).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Douglas R. Martin.

Electronic Supplementary Material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Martin, D.R., Cox, N.R., Morrison, N.E. et al. Mutation of the GM2 activator protein in a feline model of GM2 gangliosidosis. Acta Neuropathol 110, 443–450 (2005). https://doi.org/10.1007/s00401-005-1040-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00401-005-1040-6

Keywords

Navigation