Skip to main content
SpringerLink
Log in

Hexosaminidase assays

  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

β-Hexosaminidases (EC 3.2.1.52) are lysosomal enzymes that remove terminal β-glycosidically bound N-acetylglucosamine and N-acetylgalactosamine residues from a number of glycoconjugates. Reliable assay systems are particularly important for the diagnosis of a family of lysosomal storage disorders, the GM2 gangliosidoses that result from inherited β-hexosaminidase deficiency. More recently, aberrant hexosaminidase levels have also been found to be associated with a variety of inflammatory diseases. Apart from patient testing and carrier screening, practical in vitro assays are indispensable for the characterization of knock-out mice with potentially altered hexosaminidase activities, for detailed structure-function studies aimed at elucidating the enzymatic mechanism, and to characterize newly described enzyme variants from other organisms. The purpose of this article is to discuss convenient hexosaminidase assay procedures for these and other applications, using fluorogenic or chromogenic artificial substrates as well as the physiological glycolipid substrate GM2. Attempts are also made to provide an overview of less commonly used alternative techniques and to introduce recent developments enabling high-throughput screening for enzyme inhibitors.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

Notes

  1. For comprehensive advice in performing the assay for diagnosis work and for calibration and testing of procedures employed in individual laboratories, we would like to refer the reader to The International Laboratory Quality Control Carrier Testing Tay-Sachs Program of the National Tay-Sachs & Allied Disease Association (NTSAD), www.ntsad.org.

Abbreviations

BSA:

bovine serum albumin

GM2:

ganglioside GM2

GM2AP:

GM2-activator protein

GSLs:

glycosphingolipids

HexA:

β-hexosaminidase A

HexB:

β-hexosaminidase B

HexS:

β-hexosaminidase S

HAS:

human serum albumin

LUVs:

large unilamellar vesicles

MUG:

4-methylumbelliferyl-2-acetamido-2-deoxy-β-D-glucopyranoside

MUGS:

4-methylumbelliferyl-2-acetamido-2-deoxy-6-sulfo-β-D-glucopyranoside

TLC:

thin-layer chromatography

TSD:

Tay-Sachs disease

References

  1. Gravel, R.A., Kaback, M., Proia, R.L., Sandhoff, K., Suzuki, K., Suzuki, K.: The GM2 Gangliosidoses. In: Scriver, C.R., Beaudet, A.L., Sly, W., Valle, D. (eds.) The Metabolic and Molecular Bases of Inherited Disease, pp. 3827–3876. Mc Graw-Hill, New York (2001)

    Google Scholar 

  2. Watanabe, K.: Biochemical studies on carbohydrates: XXII. On animal β-N-Monoacetylglucosaminidase. J. Biochem. (Tokyo) 24, 297–303 (1936)

    CAS  Google Scholar 

  3. Sandhoff, K., Wassle, W.: Enrichment and characterization of 2 forms of human N-acetyl-beta-d-hexosaminidase. Hoppe Seylers Z. Physiol. Chem. 352, 1119–1133 (1971)

    CAS  PubMed  Google Scholar 

  4. Kolter, T., Sandhoff, K.: Sphingolipids—their metabolic pathways and the pathobiochemistry of neurodegenerative diseases. Angew. Chem. Int. Ed. Engl. 38, 1532–1568 (1999)

    Article  CAS  Google Scholar 

  5. Kolter, T., Sandhoff, K.: Sphingolipid metabolism diseases. Biochim. Biophys. Acta 1758, 2057–2079 (2006)

    Article  CAS  PubMed  Google Scholar 

  6. Proia, R.L.: Gene encoding the human beta-hexosaminidase beta chain: extensive homology of intron placement in the alpha-and beta chain genes. Proc. Natl. Acad. Sci. U.S.A. 85, 1883–1887 (1988)

    Article  CAS  PubMed  Google Scholar 

  7. Kytzia, H.J., Sandhoff, K.: Evidence for two different active sites on human beta-hexosaminidase A. Interaction of GM2 activator protein with beta-hexosaminidase A. J. Biol. Chem. 260, 7568–7572 (1985)

    CAS  PubMed  Google Scholar 

  8. Maier, T., Strater, N., Schuette, C.G., Klingenstein, R., Sandoff, K., Saenger, W.: The X-ray crystal structure of human beta-hexosaminidase B provides new insights into Sandhoff disease. J. Mol. Biol. 328, 669–681 (2003)

    Article  CAS  PubMed  Google Scholar 

  9. Mark, B.L., Mahuran, D.J., Cherney, M.M., Zhao, D., Knapp, S., James, M.N.: Crystal structure of human beta-hexosaminidase B: understanding the molecular basis of Sandhoff and Tay-Sachs disease. J. Mol. Biol. 327, 1093–1109 (2003)

    Article  CAS  PubMed  Google Scholar 

  10. Lemieux, M.J., Mark, B.L., Cherney, M.M., Withers, S.G., Mahuran, D.J., James, M.N.: Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. J. Mol. Biol. 359, 913–929 (2006)

    Article  CAS  PubMed  Google Scholar 

  11. Conzelmann, E., Sandhoff, K.: Purification and characterization of an activator protein for the degradation of glycolipids GM2 and GA2 by hexosaminidase A. Hoppe Seylers Z. Physiol. Chem. 360, 1837–1849 (1979)

    CAS  PubMed  Google Scholar 

  12. Furst, W., Sandhoff, K.: Activator proteins and topology of lysosomal sphingolipid catabolism. Biochim. Biophys. Acta 1126, 1–16 (1992)

    CAS  PubMed  Google Scholar 

  13. Li, Y.T., Li, S.C., Hasegawa, A., Ishida, H., Kiso, M., Bernardi, A., Brocca, P., Raimondi, L., Sonnino, S.: Structural basis for the resistance of Tay-Sachs ganglioside GM2 to enzymatic degradation. J. Biol. Chem. 274, 10014–10018 (1999)

    Article  CAS  PubMed  Google Scholar 

  14. Yadao, F., Hechtman, P., Kaplan, F.: Formation of a ternary complex between GM2 activator protein, GM2 ganglioside and hexosaminidase A. Biochim. Biophys. Acta 1340, 45–52 (1997)

    CAS  PubMed  Google Scholar 

  15. Xie, B., Rigat, B., Smiljanic-Georgijev, N., Deng, H., Mahuran, D.: 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 (1998)

    Article  CAS  PubMed  Google Scholar 

  16. Sandhoff, K.: The hydrolysis of Tay-Sachs ganglioside (TSG) by human N-acetyl-beta-d-hexosaminidase A. FEBS Lett. 11, 342–344 (1970)

    Article  CAS  PubMed  Google Scholar 

  17. Sandhoff, K.: Variation of beta-N-acetylhexosaminidase-pattern in Tay-Sachs disease. FEBS Lett. 4, 351–354 (1969)

    Article  CAS  PubMed  Google Scholar 

  18. Okada, S., O’Brien, J.S.: Tay-Sachs disease: generalized absence of a beta-d-N-acetylhexosaminidase component. Science 165, 698–700 (1969)

    Article  CAS  PubMed  Google Scholar 

  19. Kytzia, H.J., Hinrichs, U., Maire, I., Suzuki, K., Sandhoff, K.: Variant of GM2-gangliosidosis with hexosaminidase A having a severely changed substrate specificity. EMBO J. 2, 1201–1205 (1983)

    CAS  PubMed  Google Scholar 

  20. Conzelmann, E., Sandhoff, K.: 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. U.S.A. 75, 3979–3983 (1978)

    Article  CAS  PubMed  Google Scholar 

  21. Kolter, T., Sandhoff, K.: Principles of lysosomal membrane digestion: stimulation of sphingolipid degradation by sphingolipid activator proteins and anionic lysosomal lipids. Annu. Rev. Cell. Dev. Biol. 21, 81–103 (2005)

    Article  CAS  PubMed  Google Scholar 

  22. Conzelmann, E., Kytzia, H.J., Navon, R., Sandhoff, K.: Ganglioside GM2 N-acetyl-beta-d-galactosaminidase activity in cultured fibroblasts of late-infantile and adult GM2 gangliosidosis patients and of healthy probands with low hexosaminidase level. Am. J. Hum. Genet. 35, 900–913 (1983)

    CAS  PubMed  Google Scholar 

  23. Leinekugel, P., Michel, S., Conzelmann, E., Sandhoff, K.: Quantitative correlation between the residual activity of beta-hexosaminidase A and arylsulfatase A and the severity of the resulting lysosomal storage disease. Hum. Genet. 88, 513–523 (1992)

    Article  CAS  PubMed  Google Scholar 

  24. Klein, A., Henseler, M., Klein, C., Suzuki, K., Harzer, K., Sandhoff, K.: Sphingolipid activator protein D (sap-D) stimulates the lysosomal degradation of ceramide in vivo. Biochem. Biophys. Res. Commun. 200, 1440–1448 (1994)

    Article  CAS  PubMed  Google Scholar 

  25. Yamashita, T., Hashiramoto, A., Haluzik, M., Mizukami, H., Beck, S., Norton, A., Kono, M., Tsuji, S., Daniotti, J.L., Werth, N., Sandhoff, R., Sandhoff, K., Proia, R.L.: Enhanced insulin sensitivity in mice lacking ganglioside GM3. Proc. Natl. Acad. Sci. U.S.A. 100, 3445–3449 (2003)

    Article  CAS  PubMed  Google Scholar 

  26. Sango, K., McDonald, M.P., Crawley, J.N., Mack, M.L., Tifft, C.J., Skop, E., Starr, C.M., Hoffmann, A., Sandhoff, K., Suzuki, K., Proia, R.L.: Mice lacking both subunits of lysosomal beta-hexosaminidase display gangliosidosis and mucopolysaccharidosis. Nat. Genet. 14, 348–352 (1996)

    Article  CAS  PubMed  Google Scholar 

  27. Liu, Y., Wada, R., Kawai, H., Sango, K., Deng, C., Tai, T., McDonald, M.P., Araujo, K., Crawley, J.N., Bierfreund, U., Sandhoff, K., Suzuki, K., Proia, R.L.: A genetic model of substrate deprivation therapy for a glycosphingolipid storage disorder. J. Clin. Invest. 103, 497–505 (1999)

    Article  CAS  PubMed  Google Scholar 

  28. Tsui, Z.C., Chen, Q.R., Thomas, M.J., Samuel, M., Cui, Z.: A method for profiling gangliosides in animal tissues using electrospray ionization-tandem mass spectrometry. Anal. Biochem. 341, 251–258 (2005)

    Article  CAS  PubMed  Google Scholar 

  29. Tsuji, D., Kuroki, A., Ishibashi, Y., Itakura, T., Kuwahara, J., Yamanaka, S., Itoh, K.: Specific induction of macrophage inflammatory protein 1-alpha in glial cells of Sandhoff disease model mice associated with accumulation of N-acetylhexosaminyl glycoconjugates. J. Neurochem. 92, 1497–1507 (2005)

    Article  CAS  PubMed  Google Scholar 

  30. Tsuji, D., Higashine, Y., Matsuoka, K., Sakuraba, H., Itoh, K.: Therapeutic evaluation of GM2 gangliosidoses by ELISA using anti-GM2 ganglioside antibodies. Clin. Chim. Acta 378, 38–41 (2007)

    Article  CAS  PubMed  Google Scholar 

  31. Leaback, D.H., Walker, P.G.: Studies on glucosaminidase. 4. The fluorimetric assay of N-acetyl-beta-glucosaminidase. Biochem. J. 78, 151–156 (1961)

    CAS  PubMed  Google Scholar 

  32. Potier, M., Mameli, L., Belisle, M., Dallaire, L., Melancon, S.B.: Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-alpha-d-N-acetylneuraminate) substrate. Anal. Biochem. 94, 287–296 (1979)

    Article  CAS  PubMed  Google Scholar 

  33. Kresse, H., Fuchs, W., Glossl, J., Holtfrerich, D., Gilberg, W.: Liberation of N-acetylglucosamine-6-sulfate by human beta-N-acetylhexosaminidase A. J. Biol. Chem. 256, 12926–12932 (1981)

    CAS  PubMed  Google Scholar 

  34. Sandhoff, K., Conzelmann, E., Nehrkorn, H.: Specificity of human liver hexosaminidases A and B against glycosphingolipids GM2 and GA2. Purification of the enzymes by affinity chromatography employing specific elution. Hoppe Seylers Z. Physiol. Chem. 358, 779–787 (1977)

    CAS  PubMed  Google Scholar 

  35. Srivastava, S.K., Awasthi, Y.C., Yoshida, A., Beutler, E.: Studies on human beta-d-N-acetylhexosaminidases. I. Purification and properties. J. Biol. Chem. 249, 2043–2048 (1974)

    CAS  PubMed  Google Scholar 

  36. Suzuki, K.: Enzymatic diagnosis of sphingolipidoses. Methods Enzymol. 138, 727–762 (1987)

    Article  CAS  PubMed  Google Scholar 

  37. Hepbildikler, S.T., Sandhoff, R., Kolzer, M., Proia, R.L., Sandhoff, K.: Physiological substrates for human lysosomal beta-hexosaminidase S. J. Biol. Chem. 277, 2562–2572 (2002)

    Article  CAS  PubMed  Google Scholar 

  38. Hechtman, P., Kaplan, F.: Tay-Sachs disease screening and diagnosis: evolving technologies. DNA Cell. Biol. 12, 651–665 (1993)

    Article  CAS  PubMed  Google Scholar 

  39. Kaback, M.M.: Thermal fractionation of serum hexosaminidases: applications to heterozygote detection and diagnosis of Tay-Sach’s disease. Methods Enzymol. XXXVIII, 862–867 (1972)

    Article  Google Scholar 

  40. O’Brien, J.S., Okada, S., Chen, A., Fillerup, D.L.: Tay-Sachs disease. Detection of heterozygotes and homozygotes by serum hexosaminidase assay. N. Engl. J. Med. 283, 15–20 (1970)

    PubMed  Google Scholar 

  41. Natowicz, M.R., Prence, E.M.: Heterozygote screening for Tay-Sachs disease: past successes and future challenges. Curr. Opin. Pediatr. 8, 625–629 (1996)

    Article  CAS  PubMed  Google Scholar 

  42. Isaksson, A., Hultberg, B., Masson, P., Landels, E., Fensom, A.: Enzyme immunoassay of beta-hexosaminidase A and B in serum: carrier detection of GM2-gangliosidoses, and equivalence of enzyme activity and enzyme protein reactivity. Clin. Chem. 39, 1412–1415 (1993)

    CAS  PubMed  Google Scholar 

  43. Okada, S., Veath, M.L., Leroy, J., O’Brien, J.S.: Ganglioside GM2 storage diseases: hexosaminidase deficiencies in cultured fibroblasts. Am. J. Hum. Genet. 23, 55–61 (1971)

    CAS  PubMed  Google Scholar 

  44. Grabowski, G.A., Kruse, J.R., Goldberg, J.D., Chokkalingham, K., Gordon, R.E., Blakemore, K.J., Mahoney, M., Desnick, R.J.: First-trimester prenatal diagnosis of Tay-Sachs disease. Am. J. Hum.Genet. 36, 1369–1378 (1984)

    CAS  PubMed  Google Scholar 

  45. Callahan, J.W., Archibald, A., Skomorowski, M.A., Shuman, C., Clarke, J.T.R.: First trimester prenatal diagnosis of Tay-Sachs disease using the sulfated synthetic substrate for hexosaminidase A. Clin. Biochem. 23, 533–536 (1990)

    Article  CAS  PubMed  Google Scholar 

  46. Chamoles, N.A., Blanco, M., Gaggioli, D., Casentini, C.: Tay-Sachs and Sandhoff diseases: enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards. Clin. Chim. Acta 318, 133–137 (2002)

    Article  CAS  PubMed  Google Scholar 

  47. Lukacs, Z., Keil, A., Peters, V., Kohlschutter, A., Hoffman, G.F., Cantz, M., Kopitz, J.: Towards quality assurance in the determination of lysosomal enzymes: a two-centre study. J. Inherit. Metab. Dis. 26, 571–581 (2003)

    Article  CAS  PubMed  Google Scholar 

  48. Civallero, G., Michelin, K., de Mari, J., Viapiana, M., Burin, M., Coelho, J.C., Giugliani, R.: Twelve different enzyme assays on dried-blood filter paper samples for detection of patients with selected inherited lysosomal storage diseases. Clin. Chim. Acta 372, 98–102 (2006)

    Article  CAS  PubMed  Google Scholar 

  49. Sutton, V.R.: Tay-Sachs disease. Screening and counseling families at risk for metabolic disease. Obstet. Gynecol. Clin. North Am. 29, 287–296 (2002)

    Article  PubMed  Google Scholar 

  50. Ben-Yoseph, Y., Reid, J.E., Shapiro, B., Nadler, H.L.: Diagnosis and carrier detection of Tay-Sachs disease: direct determination of hexosaminidase A using 4-Methylumbelliferyl derivatives of b-N-Acetylglucosamine-6-Sulfate and b-N-Acetylgalactosamine-6-Sulfate. Am. J. Hum. Genet. 37, 733–748 (1985)

    CAS  PubMed  Google Scholar 

  51. Vallance, H., Morris, T.J., Coulter-Mackie, M., Lim-Steele, J., Kaback, M.: Common HEXB polymorphisms reduce serum HexA and HexB enzymatic activities, potentially masking Tay-Sachs disease carrier identification. Molec Genet Metab. 87, 122–127 (2006)

    Article  CAS  PubMed  Google Scholar 

  52. Walker, P.G., Woollen, J.W., Heyworth, R.: Studies on glucosaminidase. 5. Kidney N-acetyl-β-glucosaminidase and N-acetyl-b-galactosaminidase. Biochem. J. 79, 288–294 (1961)

    CAS  PubMed  Google Scholar 

  53. Chatterjee, S., Velicer, L.F., Sweeley, C.C.: Glycosphingolipid glycosyl hydrolases and glycosidases of synchronized human KB cells. J. Biol. Chem. 250, 4972–4979 (1975)

    CAS  PubMed  Google Scholar 

  54. Marciniak, J., Zalewska, A., Popko, J., Zwierz, K.: Optimization of an enzymatic method for the determination of lysosomal N-acetyl-beta-d-hexosaminidase and beta-glucuronidase in synovial fluid. Clin. Chem. Lab. Med. 44, 933–937 (2006)

    Article  CAS  PubMed  Google Scholar 

  55. Zwierz, K., Gindzienski, A., Glowacka, D., Porowski, T.: The degradation of glycoconjugates in the human gastric mucous membrane. Acta Med Acad Sci Hung. 38, 145–152 (1981)

    CAS  PubMed  Google Scholar 

  56. Casal, J.A., Cano, E., Tutor, J.C.: Beta-hexosaminidase isoenzyme profiles in serum, plasma, platelets and mononuclear, polymorphonuclear and unfractionated total leukocytes. Clin. Biochem. 38, 938–942 (2005)

    Article  CAS  PubMed  Google Scholar 

  57. Casal, J.A., Vizcaino, L., Garcia-Devesa, J., Tutor, J.C.: Thermodynamic study of beta-N-acetylhexosaminidase enzyme heterogeneity in human seminal plasma. Clin. Chim. Acta 355, 55–60 (2005)

    Article  CAS  PubMed  Google Scholar 

  58. Perez, L.F., Tutor, J.C.: Assay of beta-N-acetylhexosaminidase isoenzymes in different biological specimens by means of determination of their activation energies. Clin. Chem. 44, 226–231 (1998)

    CAS  PubMed  Google Scholar 

  59. Perez, L.F., Ribeiro, H.M., Casal, J.A., Pinto, R.A., Sa Miranda, M.C., Tutor, J.C.: Thermodynamic characterisation of the mutated isoenzyme A of beta-N-acetylhexosaminidase in GM2-gangliosidosis B1 variant. Clin. Chim. Acta 285, 45–51 (1999)

    Article  CAS  PubMed  Google Scholar 

  60. Tropak, M.B., Reid, S.P., Guiral, M., Withers, S.G., Mahuran, D.: Pharmacological enhancement of beta-hexosaminidase activity in fibroblasts from adult Tay-Sachs and Sandhoff Patients. J. Biol. Chem. 279, 13478–13487 (2004)

    Article  CAS  PubMed  Google Scholar 

  61. Tropak, M.B., Blanchard, J.E., Withers, S.G., Brown, E.D., Mahuran, D.: High-throughput screening for human lysosomal beta-N-Acetyl hexosaminidase inhibitors acting as pharmacological chaperones. Chem. Biol. 14, 153–164 (2007)

    Article  CAS  PubMed  Google Scholar 

  62. Tropak, M.B., Mahuran, D.: Lending a helping hand, screening chemical libraries for compounds that enhance beta-hexosaminidase A activity in GM2 gangliosidosis cells. FEBS J. 274, 4951–4961 (2007)

    Article  CAS  PubMed  Google Scholar 

  63. Shulman, M.L., Kulshin, V.A., Khorlin, A.Y.: A continuous fluorimetric assay for glycosidase activity: human N-acetyl-beta-d-hexosaminidase. Anal. Biochem. 101, 342–348 (1980)

    Article  CAS  PubMed  Google Scholar 

  64. Erzberger, A., Conzelmann, E., Sandhoff, K.: Assay of ganglioside GM2-N-acetyl-beta-d-galactosaminidase activity in human fibroblasts employing the natural activator protein-diagnosis of variant forms of GM2 gangliosidosis. Clin. Chim. Acta 108, 361–368 (1980)

    Article  CAS  PubMed  Google Scholar 

  65. O’Brien, J.S., Norden, G.W., Miller, A.L., Frost, R.G., Kelly, T.E.: Ganglioside GM2 N-acetyl-beta-d-galactosaminidase and asialo GM2 (GA2) N-acetyl-beta-d-galactosaminidase; studies in human skin fibroblasts. Clin. Genet. 11, 171–183 (1977)

    Article  PubMed  Google Scholar 

  66. Svennerholm, L., Hakansson, G., Mansson, J.E., Vanier, M.T.: The assay of sphingolipid hydrolases in white blood cells with labelled natural substrates. Clin. Chim. Acta 92, 53–64 (1979)

    Article  CAS  PubMed  Google Scholar 

  67. Klima, H., Klein, A., van Echten, G., Schwarzmann, G., Suzuki, K., Sandhoff, K.: Over-expression of a functionally active human GM2-activator protein in Escherichia coli. Biochem. J. 292, 571–576 (1993)

    CAS  PubMed  Google Scholar 

  68. Wu, Y.Y., Lockyer, J.M., Sugiyama, E., Pavlova, N.V., Li, Y.T., Li, S.C.: Expression and specificity of human GM2 activator protein. J. Biol. Chem. 269, 16276–16283 (1994)

    CAS  PubMed  Google Scholar 

  69. Wendeler, M., Lemm, T., Weisgerber, J., Hoernschemeyer, J., Bartelsen, O., Schepers, U., Sandhoff, K.: Expression of recombinant human GM2-activator protein in insect cells: purification and characterization by mass spectrometry. Protein Expr. Purif. 27, 259–266 (2003)

    Article  CAS  PubMed  Google Scholar 

  70. Wendeler, M., Hoernschemeyer, J., John, M., Werth, N., Schoeniger, M., Lemm, T., Hartmann, R., Kessler, H., Sandhoff, K.: Expression of the GM2-activator protein in the methylotrophic yeast Pichia pastoris, purification, isotopic labeling, and biophysical characterization. Protein Expr. Purif. 34, 147–157 (2004)

    Article  CAS  PubMed  Google Scholar 

  71. Werth, N., Schuette, C.G., Wilkening, G., Lemm, T., Sandhoff, K.: Degradation of membrane-bound ganglioside GM2 by beta-hexosaminidase A. Stimulation by GM2 activator protein and lysosomal lipids. J. Biol. Chem. 276, 12685–12690 (2001)

    Article  CAS  PubMed  Google Scholar 

  72. Novak, A., Lowden, J.A., Gravel, Y.L., Wolfe, L.S.: Preparation of radiolabeled GM2 and GA2 gangliosides. J. Lipid Res. 20, 678–681 (1979)

    CAS  PubMed  Google Scholar 

  73. Suzuki, Y., Suzuki, K.: Specific radioactive labeling of terminal N-acetylgalactosamine of glycosphingolipids by the galactose oxidase-sodium borohydride method. J. Lipid Res. 13, 687–690 (1972)

    CAS  PubMed  Google Scholar 

  74. Sonnino, S., Nicolini, M., Chigorno, V.: Preparation of radiolabeled gangliosides. Glycobiology 6, 479–487 (1996)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are indebted to Thomas Kolter and Hichem Gallala, Bonn, for carefully reading the manuscript and to Thomas Kolter for kindly providing Fig. 1. We thank the Deutsche Forschungsgemeinschaft for financial support (SFB 645).

Author information

Author notes

  1. Michaela Wendeler

    Present address: NIH/National Cancer Institute, Frederick, MD, 21702, USA

Authors and Affiliations

  1. LIMES, Membrane Biology and Lipid Biochemistry Unit, c/o Kekulé-Institut für Organische Chemie und Biochemie der Universität Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany

    Michaela Wendeler

  2. LIMES, c/o Kekulé-Institut für Organische Chemie und Biochemie der Universität Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany

    Konrad Sandhoff

Authors
  1. Michaela Wendeler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Konrad Sandhoff
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Konrad Sandhoff.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wendeler, M., Sandhoff, K. Hexosaminidase assays. Glycoconj J 26, 945–952 (2009). https://doi.org/10.1007/s10719-008-9137-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10719-008-9137-5

Keywords

Search

Navigation