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Several dystrophin-glycoprotein complex members are present in crude surface membranes but they are sodium dodecyl sulphate invisible in KCl-washed microsomes from mdx mouse muscle

Abstract

The dystrophin-glycoprotein complex (DGC) is a large trans-sarcolemmal complex that provides a linkage between the subsarcolemmal cytoskeleton and the extracellular matrix. In skeletal muscle, it consists of the dystroglycan, sarcoglycan and cytoplasmic complexes, with dystrophin forming the core protein. The DGC has been described as being absent or greatly reduced in dystrophin-deficient muscles, and this lack is considered to be involved in the dystrophic phenotype. Such a decrease in the DGC content was observed in dystrophin-deficient muscle from humans with muscular dystrophy and in mice with X-linked muscular dystrophy (mdx mice). These deficits were observed in total muscle homogenates and in partially membrane-purified muscle fractions, the so-called KCl-washed microsomes. Here, we report that most of the proteins of the DGC are actually present at normal levels in the mdx mouse muscle plasma membrane. The proteins are detected in dystrophic animal muscles when the immunoblot assay is performed with crude surface membrane fractions instead of the usually employed KCl-washed microsomes. We propose that these proteins form SDS-insoluble membrane complexes when dystrophin is absent.

Abbreviations

AU:

arbitrary unit

CSM:

crude surface membranes

DGC:

dystrophin-glycoprotein complex

DMD:

Duchenne muscular dystrophy

KCl-Ms:

KCl-washed microsomes

mdx :

X-linked muscular dystrophy

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

References

  1. Hoffman, E.P., Brown, R.H. and Kunkel, L.M. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 51 (1987) 919–928.

    Article  PubMed  CAS  Google Scholar 

  2. Ohlendieck, K., Ervasti, J.M., Snook, J.B. and Campbell, K.P. Dystrophinglycoprotein complex is highly enriched in isolated skeletal muscle sarcolemma. J. Cell Biol. 112 (1991) 135–148.

    Article  PubMed  CAS  Google Scholar 

  3. Ervasti, J., Ohlendieck, K., Kahl, S., Gaver, M. and Campbell, K. Deficiency of a glycoprotein component of the dystrophin complex in dystrophic muscle. Nature 345 (1990) 315–319.

    Article  PubMed  CAS  Google Scholar 

  4. Campbell, K. and Kahl, S. Association of dystrophin and an integral membrane glycoprotein. Nature 338 (1989) 259–262.

    Article  PubMed  CAS  Google Scholar 

  5. Ervasti, J.M. and Sonnemann, K. Biology of the striated muscle dystrophin-glycoprotein complex. Int. Rev. Cytol. 265 (2008) 191–225.

    Article  PubMed  CAS  Google Scholar 

  6. Ibraghimov-Beskrovnaya, O., Ervasti, J., Leveille, C., Slaughter, C., Sernett, S. and Campbell, K. Primary structure of dystrophin-associated glycoproteins linking dystrophin to the extracellular matrix. Nature 355 (1992) 696–702.

    Article  PubMed  CAS  Google Scholar 

  7. Smalheiser, N.R. and Kim, E. Purification of cranin, a laminin binding membrane protein. Identity with dystroglycan and reassessment of its carbohydrate moieties. J. Biol. Chem. 270 (1995) 15425–15433.

    Article  PubMed  CAS  Google Scholar 

  8. Winder, S. The complexities of dystroglycan. Trends Biochem. Sci. 26 (2001) 118–124.

    Article  PubMed  CAS  Google Scholar 

  9. Chung, W. and Campanelli, J.T. WW and EF hand domains of dystrophinfamily proteins mediate dystroglycan binding. Mol. Cell Biol. Res. Commun. 2 (1999) 162–171.

    Article  PubMed  CAS  Google Scholar 

  10. Huang, X., Poy, F., Zhang, R., Joachimiak, A., Sudol, M. and Eck, M.J. Structure of a WW domain containing fragment of dystrophin in complex with beta-dystroglycan. Nat. Struct. Biol. 7 (2000) 634–638.

    Article  PubMed  CAS  Google Scholar 

  11. Yang, B., Jung, D., Motto, D., Meyer, J., Koretzky, G. and Campbell, K.P. SH3 Domain-mediated Interaction of Dystroglycan and Grb2. J. Biol. Chem. 270 (1995) 11711–11714.

    Article  PubMed  CAS  Google Scholar 

  12. Hnia, K., Zouiten, D., Cantel, S., Chazalette, D., Hugon, G., Fehrentz, J.A., Masmoudi, A., Diment, A., Bramham, J., Mornet, D. and Winder, S.J. ZZ domain of dystrophin and utrophin: topology and mapping of a betadystroglycan interaction site. Biochem. J. 401 (2007) 667–677.

    Article  PubMed  CAS  Google Scholar 

  13. Yoshida, T., Pan, Y., Hanada, H., Iwata, Y. and Shigekawa, M. Bidirectional signaling between sarcoglycans and the integrin adhesion system in cultured L6 myocytes. J. Biol. Chem. 273 (1998) 1583–1590.

    Article  PubMed  CAS  Google Scholar 

  14. Crosbie, R.H., Lebakken, C.S., Holt, K.H., Venzke, D.P., Straub, V., Lee, J.C., Grady, R.M., Chamberlain, J.S., Sanes, J.R. and Campbell, K.P. Membrane targeting and stabilization of sarcospan is mediated by the sarcoglycan subcomplex. J. Cell Biol. 145 (1999) 153–165.

    Article  PubMed  CAS  Google Scholar 

  15. Yoshida, M., Hama, H., Ishikawa-Sakurai, M., Imamura, M., Mizuno, Y., Araishi, K., Wakabayashi-Takai, E., Noguchi, S., Sasaoka, T. and Ozawa, E. Biochemical evidence for association of dystrobrevin with the sarcoglycan-sarcospan complex as a basis for understanding sarcoglycanopathy. Hum. Mol. Genet. 9 (2000) 1033–1040.

    Article  PubMed  CAS  Google Scholar 

  16. Ahn, A.H., Freener, C.A., Gussoni, E., Yoshida, M., Ozawa, E. and Kunkel, L.M. The three human syntrophin genes are expressed in diverse tissues, have distinct chromosomal locations, and each bind to dystrophin and its relatives. J. Biol. Chem. 271 (1996) 2724–2730.

    Article  PubMed  CAS  Google Scholar 

  17. Peters, M.F., Adams, M.E. and Froehner, S.C. Differential association of syntrophin pairs with the dystrophin complex. J.Cell Biol. 138 (1997) 81–93.

    Article  PubMed  CAS  Google Scholar 

  18. Ervasti, J.M. Dystrophin, its interactions with other proteins, and implications for muscular dystrophy. Biochim. Biophys. Acta 1772 (2007) 108–117.

    PubMed  CAS  Google Scholar 

  19. Peters, M., O’Brien, K., Sadoulet-puccio, H., Kunkel, L., Adam, M. and Froehner, S. β-dystrobrevin,a new member of the dystrophin family. J. Biol. Chem. 272 (1997) 31561–31569.

    Article  PubMed  CAS  Google Scholar 

  20. Blake, D.J. Dystrobrevin dynamics in muscle-cell signalling: a possible target for therapeutic intervention in Duchenne muscular dystrophy? Neuromuscul. Disord. 12Suppl 1 (2002) S110–117.

    Article  PubMed  Google Scholar 

  21. Campbell, K.P. Three muscular dystrophies: loss of cytoskeleton-extracellular matrix linkage. Cell 80 (1995) 675–679.

    Article  PubMed  CAS  Google Scholar 

  22. Petrof, B.J., Shrager, J.B., Stedmann, H.H., Kelly, A.M. and Sweeney, H.L. Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc. Natl. Acad. Sci. USA 90 (1993) 3710–3714.

    Article  PubMed  CAS  Google Scholar 

  23. Ohlendieck, K. and Campbell, K.P. Dystrophin-associated proteins are greatly reduced in skeletal muscle from mdx mice. J. Cell Biol. 115 (1991) 1685–1694.

    Article  PubMed  CAS  Google Scholar 

  24. Ohlendieck, K., Matsumura, P., Ionasescu, V., Towbin, J., Bosch, E., Weinstein, S., Sernett, S. and Campbell, K.P. Duchenne muscular dystrophy: deficiency of dystrophin-associated proteins in the sarcolemma. Neurology 43 (1993) 795–800.

    PubMed  CAS  Google Scholar 

  25. Ozawa, E., Yoshida, M., Suzuki, A., Mizuno, Y., Hagiwara, Y. and Noguchi, S. Dystrophin-associated proteins in muscular dystrophy. Hum. Mol. Genet. 4Spec No (1995) 1711–1716.

    PubMed  CAS  Google Scholar 

  26. Hack, A., Lam, M., Cordier, L., Shoturma, D.I., Ly, C.T., Hadhazy, M.A., Hadhazy, M.R., Sweeney, H.L. and McNally, E.M. Differential requirement for individual sarcoglycans and dystrophin in the assembly and function of the dystrophin-glycoprotein complex. J. Cell Sci. 113 (2000) 2535–2544.

    PubMed  CAS  Google Scholar 

  27. Rouger, K., Cunff, M.L., Steenman, M., Potier, M., Gibelin, N., Dechene, C. and Leger, J. Global/temporal gene expression in diaphragm and hindlimb muscles of dystrophin-deficient (mdx) mice. Am. J. Physiol. Cell Physiol. 253 (2002) C773–C784.

    Google Scholar 

  28. Cluchague, N., Moreau, C., Rocher, C., Pottier, S., Leray, G., Chérel, Y. and Le Rumeur, E. β-dystroglycan can be revealed in microsomes from mdx mouse muscle by detergent treatment. FEBS Letters 572 (2004) 216–220.

    Article  PubMed  CAS  Google Scholar 

  29. Ohlendieck, K. Characterisation of the dystrophin-related protein utrophin in highly purified skeletal muscle sarcolemma vesicles. Biochim. Biophys. Acta 1283 (1996) 215–222.

    Article  PubMed  Google Scholar 

  30. Laemmli, U. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 (1970) 680–685.

    Article  PubMed  CAS  Google Scholar 

  31. Towbin, H., Staehelin, T. and Gordon, J. Electrophoresis transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76 (1979) 4350–4354.

    Article  PubMed  CAS  Google Scholar 

  32. Cox, G.A., Sunada, Y., Campbell, K.P. and Chamberlain, J.S. Dp71 can restore the dystrophin-associated glycoprotein complex in muscle but fails to prevent dystrophy, Nature Genet. 8 (1994) 333–339.

    Article  PubMed  CAS  Google Scholar 

  33. Wells, D.J., Wells, K.E., Asante, E.A., Turner, G., Sunada, Y., Campbell, K.P., Walsh, F.S. and Dickson, G. Expression of human full-length and minidystrophin in transgenic mdx mice: implications for gene therapy of Duchenne muscular dystrophy. Hum. Mol. Genet. 4 (1995) 1245–1250.

    Article  PubMed  CAS  Google Scholar 

  34. Matsumura, K., Tome, F.M., Collin, H., Leturcq, F., Jeanpierre, M., Kaplan, J.C., Fardeau, M. and Campbell, K.P. Expression of dystrophinassociated proteins in dystrophin-positive muscle fibers (revertants) in Duchenne muscular dystrophy. Neuromuscul. Disord. 4 (1994) 115–120.

    Article  PubMed  CAS  Google Scholar 

  35. Rafael, J.A., Sunada, Y., Cole, N.M., Campbell, K.P., Faulkner, J.A. and Chamberlain, J.S. Prevention of dystrophic pathology in mdx mice by a truncated dystrophin isoform. Hum. Mol. Genet. 3 (1994) 1725–1733.

    Article  PubMed  CAS  Google Scholar 

  36. Mizuno, Y., Yoshida, M., Nonaka, I., Hirai, S. and Ozawa, E. Expression of utrophin (dystrophin-related protein) and dystrophin-associated glycoproteins in muscles from patients with Duchenne muscular dystrophy. Muscle Nerve 17 (1994) 206–216.

    Article  PubMed  CAS  Google Scholar 

  37. Crawford, G.E., Faulkner, J.A., Crosbie, R.H., Campbell, K.P., Froehner, S.C. and Chaberlain, J.S. Assembly of the Dystrophin- associated Protein Complex Does Not Require the Dystrophin COOH-terminal Domain. J. Cell Biol. 150 (2000) 1399–1409.

    Article  PubMed  CAS  Google Scholar 

  38. DelloRusso, C., Scott, J., Hartigan-O’Connor, D., Salvatori, G., Barjot, C., Robinson, A., Crawford, R., Brooks, S. and Chamberlain, J.S. Functional correction of adult mdx mouse muscle using gutted adenoviral vectors expressing full-length dystrophin. Proc. Natl. Acad. Sci. USA 99 (2002) 12979–12984.

    Article  PubMed  CAS  Google Scholar 

  39. Harper, S.Q., Hauser, M.A., DelloRusso, C., Duan, D., Crawford, R.W., Phelps, S.F., Harper, H.A., Robinson, A.S., Engelhardt, J.F., Brooks, S.V. and Chamberlain, J.S. Modular flexibility of dystrophin: implications for gene therapy of Duchenne muscular dystrophy. Nature Med. 8 (2002) 253–261.

    Article  PubMed  CAS  Google Scholar 

  40. Gardner, K., Kearney, J., Edwards, J. and Rafael-Fortney, J. Restoration of all dystrophin protein interactions by functional domains in trans does not rescue dystrophy. Gene therapy 13 (2005) 744–751.

    Article  CAS  Google Scholar 

  41. Ervasti, J. and Campbell, K. A Role for the Dystrophin-Glycoprotein Complex as a Transmembrane Linker between Laminin and Actin. J. Cell Biol. 122 (1993) 809–823.

    Article  PubMed  CAS  Google Scholar 

  42. Rouger, K., Brault, M., Daval, N., Leroux, I., Guigand, L., Lesoeur, J. and Chérel, Y. Muscle satellite cell heterogeneity: in vitro and in vivo evidences for populations that fuse differently. Cell Tissue Res. 317 (2004) 319–326.

    Article  PubMed  Google Scholar 

  43. Yamada, H., Saito, F., Fukuta-Ohi, H., Zhong, D., Hase, A., Arai, K., Okuyama, A., Maekawa, R., Shimizu, T. and Matsumura K. Processing of β-dystroglycan by matrix metalloproteinase disrupts the link between the extracellular matrix and cell membrane via the dystroglycan complex. Hum. Mol. Genet. 10 (2001) 1563–1569.

    Article  PubMed  CAS  Google Scholar 

  44. Armstrong, S., Latham, C. and Ganote, C. An ischemic beta-dystroglycan degradation product: correlation with irreversible injury in adult rabbit cardiomyocytes. Mol. Cell. Biochem. 242 (2003) 71–79.

    Article  PubMed  CAS  Google Scholar 

  45. Bierczynska-Krzysik, A., Kang, S.U., Silberrring, J. and Lubec, G. Mass spectrometrical identification of brain proteins including highly insoluble and transmembrane proteins. Neurochem. Int. 49 (2006) 245–255.

    Article  PubMed  CAS  Google Scholar 

  46. Zhou, J., Zhou, T., Cao, R., Liu, Z., Shen, J., Chen, P., Wang, X. and Liang, S. Evaluation of the application of sodium deoxycholate to proteomic analysis of rat hippocampal plasma membrane. J. Proteome Res. 5 (2006) 2547–2553.

    Article  PubMed  CAS  Google Scholar 

  47. Galvin, J.E., Palamand, D., Strider, J., Milone, M. and Pestronk, A. The muscle protein dysferlin accumulates in the Alzheimer brain. Acta Neuropathol. 112 (2006) 665–671.

    Article  PubMed  CAS  Google Scholar 

  48. Wong, S.L., Chan, W.M. and Chan, H. Y. Sodium dodecyl sulfateinsoluble oligomers are involved in polyglutamine degeneration. FASEB J. (2008)

  49. Cappai, R., Leck, S.L., Tew, D.J., Williamson, N.A., Smith, D.P., Galatis, D., Sharples, R.A., Curtain, C.C., Ali, F.E., Cherny, R.A., Culvenor, J.G., Bottomley, S.P., Masters, C.L., Barnham, K.J. and Hill, A.F. Dopamine promotes alpha-synuclein aggregation into SDS-resistant soluble oligomers via a distinct folding pathway. FASEB J. 19 (2005) 1377–1379.

    PubMed  CAS  Google Scholar 

  50. Greenberg, D.S., Sunada, Y., Campbell, K.P., Yaffe, D. and Nudel, U. Exogenous Dp71 restores the levels of dystrophin-associated proteins but does not alleviate muscle damage in mdx mice. Nature Genet. 8 (1994) 340–344.

    Article  PubMed  CAS  Google Scholar 

  51. Rezniczek, G.A., Konieczny, P., Nikolic, B., Reipert, S., Schneller, D., Abrahamsberg, C., Davies, K.E., Winder, S.J. and Wiche, G. Plectin 1f scaffolding at the sarcolemma of dystrophic (mdx) muscle fibers through multiple interactions with beta-dystroglycan. J. Cell Biol. 176 (2007) 965–977.

    Article  PubMed  CAS  Google Scholar 

  52. Batchelor, C.L. and Winder, S.J. Sparks, signals and shock absorbers: how dystrophin loss causes muscular dystrophy. Trends Cell Biol. 16 (2006) 198–205.

    Article  PubMed  CAS  Google Scholar 

  53. Hack, A., Groth, M. and McNally, E.M. Sarcoglycans in muscular dystrophy. Microsc. Res. Tech. 48 (2000) 167–180.

    Article  PubMed  CAS  Google Scholar 

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Daval, S., Rocher, C., Cherel, Y. et al. Several dystrophin-glycoprotein complex members are present in crude surface membranes but they are sodium dodecyl sulphate invisible in KCl-washed microsomes from mdx mouse muscle. Cell Mol Biol Lett 15, 134–152 (2010). https://doi.org/10.2478/s11658-009-0039-8

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