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Laboratory Guide to the Methods in Biochemical Genetics pp 287–324Cite as

Mucopolysaccharides

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Abstract

Mucopolysaccharides or glycosaminoglycans (GAGs) are polysaccharides, which consist of repeating disaccharide units. Deficiencies of certain lysosomal enzymes result in accumulation of GAGs in tissue and urine. All mucopolysaccharidoses (MPS) represent chronic, progressive disorders which exhibit a wide variety of clinical manifestations, from skeletal abnormalities to intellectual impairment. Diagnosis should be based on firm clinical observations before biochemical tests are initiated. A four-step process is then recommended. The first step should be a relatively unspecific screening test for urinary GAG excretion. The second step should be the differentiation of urinary GAGs by electrophoresis or thin-layer chromatography (TLC). In the third step the enzyme deficiency should be demonstrated. Finally, for future genetic counseling, the identification of the distinct mutations in both alleles should be considered. In this chapter, the dimethylene method as a urinary screening test, electrophoresis and TLC for GAG differentiation and the enzyme assays from leukocytes as well as dried blood will be introduced. Furthermore, pitfalls will be discussed and a short survey of alternative methods will be given.

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References

  1. Baum H, Dodgson KS, Spencer B (1959) The assay of arylsulphatase A and B in human urine. Clin Chim Acta 4:453–455

    Article  PubMed  CAS  Google Scholar 

  2. Berman ER, Vered J, Bach G (1971) A reliable spot test for mucopolysaccharidoses. Clin Chem 17:886–890

    PubMed  CAS  Google Scholar 

  3. Berry HK, Berry MA, Spinanger J (1960) A paper spot test useful in study of Hurler’s syndrome. J Lab Clin Med 55:136–138

    PubMed  CAS  Google Scholar 

  4. Berry HK (1987) Screening for mucopolysaccharide disorders with the Berry spot test. Clin Biochem 20:365–371

    Article  PubMed  CAS  Google Scholar 

  5. Bitter T, Muir HM (1962) A modified uronic acid carbazole reaction. Anal Biochem 4:330–334

    Article  PubMed  CAS  Google Scholar 

  6. Brante G (1952) Gargoylism: a mucopolysaccharidosis. Scand J Clin Lab Invest 4:43–46

    Article  PubMed  CAS  Google Scholar 

  7. Brimble A, Pennock C, Stone J (1990) Results of a quality assurance exercise for urinary glycosaminoglycan excretion. Ann Clin Biochem 27:133–138

    PubMed  Google Scholar 

  8. Byers S, Rozaklis T, Brumfield LK, Ranieri E, Hopwood JJ (1998) Glycosaminoglycan accumulation and excretion in the mucopolysaccharidoses: characterization and basis of a diagnostic test for MPS. Mol Genet Metab 65:282–290

    Article  PubMed  CAS  Google Scholar 

  9. Cappelletti R, del Rosso M, Chiarugi VP (1979) A New electrophoretic method for the complete separation of all known animal glycosaminoglycans in a monodimensional run. Anal Biochem 99:311–315

    Article  PubMed  CAS  Google Scholar 

  10. Carson NAJ, Neill DW (1962) Metabolic abnormalities detected in a survey of mentally backward individuals in Northern Ireland. Arch Dis Child 37:505–513

    PubMed  CAS  Google Scholar 

  11. Chamoles NA, Blanco MB, Gaggioli D, Casentini C (2001) Hurler-like Phenotype: enzymatic diagnosis in dried blood spots on filter paper. Clin Chem 47:2098–2102

    PubMed  CAS  Google Scholar 

  12. Chamoles NA, Blanco M, Gaggioli D (2001) Diagnosis of α-L-iduronidase deficiency in dried blood spots on filter paper: the possibility of newborn diagnosis. Clin Chem 47:780–781

    PubMed  CAS  Google Scholar 

  13. Chih-Kuang C, Shuan-Pei L, Shyue-Jye L, Tuen-Jen W (2002) MPS screening methods, the Berry Spot and acid turbidity tests, cause a high incidence of false-negative results in Sanfilippo and Morquio syndromes. J Clin Lab Anal 16:253–258

    Article  PubMed  Google Scholar 

  14. De Jong JGN, Wevers RA, Laarakkers C, Poorthuis BJHM (1989) Dimethylene blue-based spectrophotometry of glycosaminoglycans in untreated urine: a rapid screening procedure for mucopolysaccharidoses. Clin Chem 35:1472–1477

    PubMed  Google Scholar 

  15. De Jong JGN, Hasselman JJF, Landeghem AAJ, Vader HL, Wevers RA (1991) The spot test is not a reliable screening procedure for mucopolysaccharidoses. Clin Chem 37:572–575

    PubMed  Google Scholar 

  16. De Jong JGN, Wevers RA, Liebrand-van Sambeek R (1992) Measuring urinary glycosaminoglycans in the presence of protein: an improved screening procedure for mucopolysaccharidoses based on dimethylene blue. Clin Chem 38:803–807

    PubMed  Google Scholar 

  17. De Jong JGN, Heijs WM, Wevers RA (1994) Mucopolysaccharidoses screening: dimethylene blue versus Alcian blue. Ann Clin Biochem 31:267–271

    PubMed  Google Scholar 

  18. Dembure PP, Drumheller JE, Barr SM, Elsas LJ (1990) Selective urinary screening for mucopolysaccharidoses. Clin Biochem 23:91–96

    Article  PubMed  CAS  Google Scholar 

  19. Dembure PP, Roesel RA (1991) Screening for mucopolysaccharidoses by analysis of urinary glycosaminoglycans. In: Hommes FA (ed) Techniques in Diagnostic Human Biochemical Genetics. Wiley-Liss, New York, pp 77–86

    Google Scholar 

  20. Di Ferrante NM (1967) The measurement of urinary mucopolysaccharides. Anal Biochem 21:98–106

    Article  PubMed  Google Scholar 

  21. Dorfman A, Lorincz AE (1957) Occurrence of urinary acid mucopolysaccharides in the Hurler syndrome. 43:443–446

    Google Scholar 

  22. Frantatoni JC, Hall CW, Neufeld EF (1968) The defect in Hurler’s and Hunter’s syndromes: faulty degradation of mucopolysaccharides. Proc Natl Acad Sci U S A 60:699–706

    Article  Google Scholar 

  23. Gässler N, Reißner C, Janzen N, Kähnert H, Kleesiek K (1993) A high-performance liquid chromatography method for the determination of glycosaminoglycans in human blood. Eur J Clin Chem Clin Biochem 31:503–511

    PubMed  Google Scholar 

  24. Glaser JH, Sly WS (1973) β-Glucuronidase deficiency mucopolysaccharidosis: methods for enzymatic diagnosis. J Lab Clin Med 82:969–977

    PubMed  CAS  Google Scholar 

  25. Gold EW (1979) A simple spectrophotometric method for estimating glycosaminoglycan concentrations. Anal Biochem 99:183–188

    Article  PubMed  CAS  Google Scholar 

  26. Gold EW (1981) The quantitative spectrophotometric estimation of total sulfated glycosaminoglycan levels. Biochim Biophys Acta 673:40–415

    Google Scholar 

  27. Hall CW, Liebaers I, Di Natale P, Neufeld EF (1978) Enzymic diagnosis of the genetic mucopolysaccharide storage disorders. Methods Enzymol 50:439–456

    PubMed  CAS  Google Scholar 

  28. Hata R, Nagai Y (1972) A rapid and micro method for separation of acidic glycosaminoglycans by two-dimensional electrophoresis. Anal Biochem 45:462–468

    Article  PubMed  CAS  Google Scholar 

  29. He W, Voznyi YV, Boer AM, Kleijer WJ, van Diggelen OP (1993) A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease type D (MPS IIID). J Inherit Metab Dis 16:935–941

    Article  PubMed  CAS  Google Scholar 

  30. Hein LK, Meikle PJ, Dean CJ, Bockmann MR, Auclair D, Hopwood JJ, Brooks DA (2005) Development of an assay for the detection of mucopolysaccharidosis type VI patients using dried blood spots. Clin Chim Acta 353:67–74

    Article  PubMed  CAS  Google Scholar 

  31. Hochuli M, Wüthrich K, Steinmann B (2003) Two-dimensional NMR spectroscopy of urinary glycosaminoglycans from patients with different mucopolysaccharidoses. NMR Biomed 16:224–236

    Article  PubMed  CAS  Google Scholar 

  32. Hopwood JJ, Harrison JR (1982) High-resolution electrophoresis of urinary glycosaminoglycans: an improved screening test for the mucopolysaccharidoses. Anal Biochem 119:120–127

    Article  PubMed  CAS  Google Scholar 

  33. Hsu D, Hoffman P, Mashburn TA (1972) Micro colorimetric method of dye staining and its application to cellulose polyacetate strip electrophoresis for the determination of acid glycosaminoglycans. Anal Biochem 46:156–163

    Article  PubMed  CAS  Google Scholar 

  34. Huang K, Sukegawa K, Orii T (1985) Screening test for urinary glycosaminoglycans and differentiation of various mucopolysaccharidoses. Clin Chim Acta 151:147–156

    Article  PubMed  CAS  Google Scholar 

  35. Humbel R, Chamoles NA (1972) Sequential thin layer chromatography of urinary acid glycosaminoglycans. Clin Chim Acta 40:290–293

    Article  PubMed  CAS  Google Scholar 

  36. Iwata S, Sukegawa K, Sasaki T, Kokuryu M, Yamasita S, Noma A, Iwasa S, Kondo N, Orii T (1997) Mass screening test for mucopolysaccharidoses using the 1,9-dimethylene blue method: positive interference from paper diapers. Clin Chim Acta 264:245–250

    Article  PubMed  CAS  Google Scholar 

  37. Kaplan D (1969) Classification of the mucopolysaccharidoses based on the pattern of mucopolysacchariduria. Am J Med 47:721–729

    Article  PubMed  CAS  Google Scholar 

  38. Karpova EA, Voznyi YV, Keulemans JLM, Hoogeveen AT, Winchester B, Tsvetkova IV, van Diggelen OP (1996) A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease type A (MPS IIIA). J Inherit Metab Dis 19:278–285

    Article  PubMed  CAS  Google Scholar 

  39. Kodama C (1988) Methods for analysis of urinary glycosaminoglycans. J Chromatogr 429:293–313

    Article  PubMed  CAS  Google Scholar 

  40. Kresse H, Cantz M, von Figura K, Glössl J, Paschke E (1981) The mucopolysaccharidoses: biochemistry and clinical symptoms. Klin Wochenschr 59:867–876

    Article  PubMed  CAS  Google Scholar 

  41. Kresse H, von Figura K, Klein U, Glössl J, Paschke E, Pohlmann R (1982) Enzymic diagnosis of genetic mucopolysaccharide storage disorders. Methods Enzymol 83:559–572

    PubMed  CAS  Google Scholar 

  42. Lippiello L, Mankin HJ (1971) Thin-layer chromatographic separation of the isomeric chondroitin sulfates, dermatan sulfate and keratan sulfate. Anal Biochem 39:54–58

    Article  PubMed  CAS  Google Scholar 

  43. Lowry OH, Roseborough NJ, Farr AL, Randall RJ (1951) Protein measurements with Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  44. Lukacs Z, Santavuori P, Keil A, Steinfeld R, Kohlschütter A (2003) Rapid and simple assay for the determination of tripeptidyl peptidase and palmitoyl protein thioesterase activities in dried blood spots. Clin Chem 49:509–511

    Article  PubMed  CAS  Google Scholar 

  45. Mabe P, Valiente A, Soto V, Cornejo V, Raimann E (2004) Evaluation of reliability for urine mucopolysaccharidosis screening by dimethylene blue and Berry spot tests. Clin Chim Acta 345:135–140

    Article  PubMed  CAS  Google Scholar 

  46. Mao W, Thanawiroon C, Linhardt RJ (2002) Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan-derived oligosaccharides. Biomed Chromatogr 16:77–94

    Article  PubMed  CAS  Google Scholar 

  47. Matsuno Y, Kinoshita M, Kakehi K (2005) Fast analysis of glycosaminoglycans by microchip electrophoresis with in situ fluorescent detection using ethidium bromide. J Pharm Biomed Anal 37:429–436

    Article  PubMed  CAS  Google Scholar 

  48. McKusick VA (1969) The nosology of the mucopolysaccharidoses. Am J Med 47:730–747

    Article  PubMed  CAS  Google Scholar 

  49. Miura RO, Yamagata S, Miura Y, Harada T, Yamagata T (1995) Analysis of glycosaminoglycan-degrading enzymes by substrate gel electrophoresis (zymography) Anal Biochem 225:333–340

    Article  PubMed  CAS  Google Scholar 

  50. Narita H, Takeda Y, Takagaki K, Nakamura T, Harata S, Endo M (1995) Identification of glycosaminoglycans using high-performance liquid chromatography on a hydroxyapatite column. Anal Biochem 232:133–136

    Article  PubMed  CAS  Google Scholar 

  51. Neufeld EF, Muenzer J (2001) The mucopolysaccharidoses. In: Scriver CR, Sly WS, Childs B, Beaudet AL, Valle D, Kinzler KW, Vogelstein B (eds) The Metabolic and Molecular Basis of Inherited Disease, 8th edn. McGraw-Hill, New York, pp 3421–3452

    Google Scholar 

  52. Northover H, Cowie RA, Wraith JE (1996) Mucopolysaccharidosis type IVA (Morquio syndrome): a clinical review. J Inherit Metab Dis 19:357–365

    Article  PubMed  CAS  Google Scholar 

  53. Panin G, Naia S, Dall´Amico R, Chiandetti L, Zachello F, Catassi C, Felici, L, Coppa GV (1986) Simple spectrophotometric quantification of urinary, excretion of glycosaminoglycan sulfates. Clin Chem 32:2073–2076

    PubMed  CAS  Google Scholar 

  54. Pennock CA (1976) A review and selection of simple laboratory methods used for the study of glycosaminoglycan excretion and the diagnosis of the mucopolysaccharidoses. J Clin Pathol 29:111–123

    Article  PubMed  CAS  Google Scholar 

  55. Piraud M, Maire I, Mathieu M (1993) Pitfalls of screening for mucopolysaccharidoses by the dimethylene blue test. Clin Chem 39:163–164

    PubMed  CAS  Google Scholar 

  56. Piraud M, Boyer M, Mathieu M, Maire I (1993) Diagnosis of mucopolysaccharidoses in a clinically selected population by urinary glycosaminoglycan analysis: a study of 2000 urine samples. Clin Chim Acta 221:171–181

    Article  PubMed  CAS  Google Scholar 

  57. Proudfoot AEI, Handel TM, Johnson Z, Lau EK, LiWang P, Clark-Lewis I, Borlat F, Wells TNC, Kosco-Villbois MH (2003) Glycosaminoglycan binding and oligomerization are essential for the in vivo activity of certain chemokines. Proc Natl Acad Sci U S A 100:1885–1890

    Article  PubMed  CAS  Google Scholar 

  58. Sewell AC (1988) Urinary screening for disorders of heteroglycan metabolism. Klin Wochenschr 66:48–53

    Article  PubMed  CAS  Google Scholar 

  59. Stirling JL, Robinson D, Fensom AH, Benson PF, Baker JE (1978) Fluorimetric assay for prenatal detection of Hurler and Scheie homozygotes or heterozygotes. Lancet 1:147

    Article  PubMed  CAS  Google Scholar 

  60. Stirling JL, Robinson D, Fensom AH, Benson PF, Baker JE, Button LR (1979) Prenatal diagnosis of two Hurler fetuses using an improved assay for methylumbelliferyl-α-L-iduronidase. Lancet 2:37

    Article  PubMed  CAS  Google Scholar 

  61. Stone JE, Akhtar N, Botchway S, Pennock CA (1994) Interaction of 1,9 dimethylene blue with glycosaminoglycans. Ann Clin Biochem 31:147–152

    PubMed  CAS  Google Scholar 

  62. Toida T, Shima M, Azumaya S, Maruyama T, Toyoda H, Imanari T, Linhardt RJ (1997) Detection of glycosaminoglycans as copper (II) complex in high-performance liquid chromatography. J Chromatogr A 787:266–270

    Article  PubMed  CAS  Google Scholar 

  63. Tuschl K, Gal A, Paschke E, Kircher S, Bodamer O (2004) Mucopolysaccharidosis type ii in females: case report and review of literature. Pediatr Neurol 32:270–272

    Article  Google Scholar 

  64. Van den Lest CHA, Versteeg EMM, Veerkamp JH, van Kuppevelt TH (1994) Quantification and characterization of glycosaminoglycans at nanogram level by a combined azure A-silver staining in agarose gels. Anal Biochem 221:356–361

    Article  PubMed  Google Scholar 

  65. Van Diggelen OP, Zhao H, Kleijer WJ, Janse HC, Poorthuis BJHM, van Pelt J, Kamerling JP, Galjaard H (1990) A fluorimetric enzyme assay for the diagnosis of Morquio disease type A (MPS IV A) Clin Chim Acta 187:131–140

    Article  PubMed  Google Scholar 

  66. Voznyi YV, Karpova EA, Dudukina TV, Tsvetkova IV, Boer AM, Janse HC, van Diggelen OP (1993) A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease C (MPS III C) J Inherit Metab Dis 16:465–472

    Article  PubMed  CAS  Google Scholar 

  67. Voznyi YV, Keulemans JLM, van Diggelen OP (2001) A fluorimetric assay for the diagnosis of MPS II (Hunter’s disease). J Inherit Metab Dis 24:675–680

    Article  PubMed  CAS  Google Scholar 

  68. Wang D, Eadala B, Sadilek M, Chamoles NA, Turecek F, Scott R, Gelb MH (2005) Tandem mass spectrometric analysis of dried blood spots for screening of mucopolysaccharidoses I in newborns. Clin Chem 51:898–900

    Article  PubMed  CAS  Google Scholar 

  69. Wessler E (1968) Analytical and preparative separation of acidic glycosaminoglycans by electrophoresis in barium acetate. Anal Biochem 26:439–444

    Article  PubMed  CAS  Google Scholar 

  70. Whitley CB, Ridnour MD, Draper KA, Dutton CM, Neglia JP (1989) Diagnostic test for mucopolysaccharidosis. I. Direct method for quantifying excessive urinary glycosaminoglycan excretion. Clin Chem 35:374–379

    PubMed  CAS  Google Scholar 

  71. Wraith JE (1995) The mucopolysaccharidoses: a clinical review and guide to management. Arch Dis Child 72:263–267

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pediatrics, Metabolic Laboratory, University Hospital of Hamburg, Martinistraße 52, 20246, Hamburg, Germany

    Zoltan Lukacs

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  1. Zoltan Lukacs
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Editors and Affiliations

  1. Division of Clinical Chemistry and Biochemistry, University Children’s Hospital, Steinwiesstrasse 75, 8032, Zurich, Switzerland

    Nenad Blau

  2. Academic Medical Centre, Lab. Genetic Metabolic; Diseases Fo-224, University Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, Netherlands

    Marinius Duran

  3. Dept. of Molecular and Medical Genetics, Oregon Health & Science University, 3rd Avenue 2525, 97201, Portland, USA

    K. Michael Gibson

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Lukacs, Z. (2008). Mucopolysaccharides. In: Blau, N., Duran, M., Gibson, K. (eds) Laboratory Guide to the Methods in Biochemical Genetics. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-76698-8_17

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  • DOI: https://doi.org/10.1007/978-3-540-76698-8_17

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