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Comparative determination of the particle weight of glycoproteins by SDS-PAGE and analytical ultracentrifugation

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Progress in Analytical Ultracentrifugation

Part of the book series: Progress in Colloid & Polymer Science ((PROGCOLLOID,volume 86))

Abstract

The particle weight of 21 glycoproteins has been determined by SDS-PAGE and high-speed sedimentation equilibrium, focussing on the influence of carbohydrate content, partial specific volume (\(\bar v_2\)), and relative molar mass (M r) on the results of both techniques. Results were compared with literature data and with 22 nonconjugated proteins yielding the following results: (i) \(\bar v_2\)of native glycoproteins decreases linearly with increasing carbohydrate content. Therefore, \(\bar v_2 - values\)of glycoproteins not accessible to experimental determination may be calculated. (ii) The determination of M r of glycoprotein subunits by SDS-PAGE shows anomalies in the electrophoretic behaviour. Small amounts of impurities have no influence on the result, if one refers only to the major band(s). Systematic variation of the electrophoretic parameters T and C, as well as use of Ferguson plots only leads to insignificant improvements. There is no correlation between the carbohydrate content and anomalies in the electrophoretic behaviour. A possible explanation for this behaviour may be found in different conformations of the sugar moiety, different interactions with the electrophoresis gel, and different extent of SDS binding, (iii) Analytical ultracentrifugation yields accurate values for M r of native glycoproteins, independent of the carbohydrate content and the size of the molecule. The application of this technique, however, requires the exact knowledge of \(\bar v_2\)and the absence of impurities, (iv) A comparison of the molar subunit masses obtained by the two methods with literature values clearly proves the superiority of analytical ultracentrifugation, when compared with SDS-PAGE.

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References

  1. Gottschalk A, ed (1972) Glycoproteins, Parts A and B, 2nd ed. Elsevier, Amsterdam-London-New York

    Google Scholar 

  2. Gibbons RA (1972) In: Gottschalk A (ed) Glycoproteins, Part A, 2nd ed. Elsevier, Amsterdam-London-New York, pp 31–140

    Google Scholar 

  3. Fish WW (1975) In: Korn ED (ed) Methods in Membrane Biology, Vol 4. Plenum Press, New York-London, pp 189–276

    Google Scholar 

  4. Beeley JG (1985) In: Burdon RH, van Knippenberg PH (eds) Laboratory Techniques in Biochemistry and Molecular Biology, Vol 16. Elsevier, Amsterdam-New York-Oxford, pp 63–99

    Google Scholar 

  5. Andrews AT (1986) Electrophoresis: Theory, Techniques, and Biochemical and Clinical Applications, 2nd ed. Clarendon Press, Oxford

    Google Scholar 

  6. Wagner H, Blasius E (eds) (1989) Praxis der elektrophoretischen Trennmethoden. Springer-Verlag, Berlin-Heidelberg-New York-London-Paris-Tokyo

    Google Scholar 

  7. Hames BD, Rickwood D (eds) (1990) Gel Electrophoresis of Proteins, A Practical Approach, 2nd ed. IRL Press at Oxford University Press, Oxford-New York-Tokyo

    Google Scholar 

  8. Schubert D (1970) J Mol Biol 51:287–301

    Article  CAS  Google Scholar 

  9. Bretscher MS (1971) Nature New Biol 231:229–232

    Article  CAS  Google Scholar 

  10. Glossmann H, Neville DM Jr (1971) J Biol Chem 246:6339–6346

    CAS  Google Scholar 

  11. Segrest JP, Jackson RL, Andrews EP, Marchesi VT (1971) Biochem Biophys Res Commun 44:390–395

    Article  CAS  Google Scholar 

  12. Segrest JP, Jackson RL (1972) Methods Enzymol 28:54–63

    Google Scholar 

  13. Banker GA, Cotman CW (1972) J Biol Chem 247:5856–5861

    CAS  Google Scholar 

  14. Westphal U, Burton RM, Harding GB (1975) Methods Enzymol 36:91–104

    CAS  Google Scholar 

  15. Leach BS, Collawn JF Jr, Fish WW (1980) Biochemistry 19:5734–5741

    Article  CAS  Google Scholar 

  16. Creeth JM, Pain RH (1967) Progr Biophys Molec Biol 17:217–287

    Article  CAS  Google Scholar 

  17. Chervenka CH (1973) A Manual of Methods for the Analytical Ultracentrifuge. Spinco Division of Beckman Instruments, Palo Alto

    Google Scholar 

  18. Van Holde KE (1975) In: Neurath H, Hill RC (eds) The Proteins, Vol 1, 3rd ed. Academic Press, New York-San Francisco-London, pp 225–291

    Google Scholar 

  19. Krischner K, Voigt B (1968) Hoppe-Seyler's Z Physiol Chem 349:632–644

    Google Scholar 

  20. Stallcup WB, Mockrin SC, Koshland DE Jr (1972) J Biol Chem 247:6277–6279

    CAS  Google Scholar 

  21. Bradford MM (1976) Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  22. Moore S (1968) J Biol Chem 243:6281–6283

    CAS  Google Scholar 

  23. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Anal Chem 28:350–356

    Article  CAS  Google Scholar 

  24. François C, Marshall RD, Neuberger A (1962) Biochem J 83:335–341

    Google Scholar 

  25. Boehringer (1987) Glycohydrolases. Boehringer, Mannheim

    Google Scholar 

  26. Kratky O, Leopold H, Stabinger H (1973) Methods Enzymol 27:98–110

    CAS  Google Scholar 

  27. Durchschlag H, Jaenicke R (1982) Biochem Biophys Res Commun 108:1074–1079

    Article  CAS  Google Scholar 

  28. Durchschlag H (1986) In: Hinz H-J (ed) Thermodynamic Data for Biochemistry and Biotechnology. Springer Verlag, Berlin-Heidelberg-New York-Tokyo, pp 45–128

    Google Scholar 

  29. Durchschlag H (1988) Biochem (Life Sci Adv) 7:181–188

    Google Scholar 

  30. Durchschlag H (1989) Colloid Polym Sci 267:1139–1150

    Article  CAS  Google Scholar 

  31. Ornstein L (1964) Ann N Y Acad Sci 121:321–349

    Article  CAS  Google Scholar 

  32. Davis BJ (1964) Ann N Y Acad Sci 121:404–427

    Article  CAS  Google Scholar 

  33. Shapiro AL, Viñuela E, Maizel JV Jr (1967) Biochem Biophys Res Commun 28:815–820

    Article  CAS  Google Scholar 

  34. Dunker AK, Rueckert RR (1969) J Biol Chem 244:5074–5080

    CAS  Google Scholar 

  35. Weber K, Osborn M (1969) J Biol Chem 244:4406–4412

    CAS  Google Scholar 

  36. Weber K, Pringle JR, Osborn M (1972) Methods Enzymol 26:3–27

    Article  CAS  Google Scholar 

  37. Kondo T, Tarutani O, Ui N (1981) J Biochem 89:379–384

    CAS  Google Scholar 

  38. Laemmli UK (1970) Nature 227:680–685

    Article  CAS  Google Scholar 

  39. Laemmli UK, Favre M (1973) J Mol Biol 80:575–599

    Article  CAS  Google Scholar 

  40. Zacharius RM, Zell TE, Morrison JH, Woodlock JJ (1969) Anal Biochem 30:148–152

    Article  CAS  Google Scholar 

  41. Colonna WJ, Cano FR, Lampen JO (1975) Biochim Biophys Acta 386:293–300

    CAS  Google Scholar 

  42. Ferguson KA (1964) Metab Clin exp 13:985–1002

    CAS  Google Scholar 

  43. Neville DM Jr (1971) J Biol Chem 246:6328–6334

    CAS  Google Scholar 

  44. Durchschlag H, Jaenicke R (1983) Int J Biol Macromol 5:143–148

    Article  CAS  Google Scholar 

  45. Yphantis DA (1964) Biochemistry 3:297–317

    Article  CAS  Google Scholar 

  46. Smith MH (1970) In: Sober HA (ed) CRC Handbook of Biochemistry, 2nd ed. The Chemical Rubber Co, Cleveland, pp C3–C35

    Google Scholar 

  47. Klotz IM, Darnall DW (1970) In: Sober HA (ed) CRC Handbook of Biochemistry, 2nd ed. The Chemical Rubber Co, Cleveland, pp C62–C66

    Google Scholar 

  48. Fasman GD (ed) (1976) CRC Handbook of Biochemistry and Molecular Biology, 3rd ed, Proteins Vol 2 and 3. CRC Press, Cleveland

    Google Scholar 

  49. Soodak M (1976) In: Fasman GD (ed) CRC Handbook of Biochemistry and Molecular Biology, 3rd ed, Proteins Vol 2. CRC Press, Cleveland, pp 257–275

    Google Scholar 

  50. Dayhoff MO (ed) (1972) Atlas of Protein Sequence and Structure, Vol 5: (1973) Vol 5 Suppl 1; (1976) Vol 5 Suppl 2; (1978) Vol 5 Suppl 3. National Biomed Research Foundation, Washington DC

    Google Scholar 

  51. Schultze HE, Heremans JF (1966) Molecular Biology of Human Proteins: With Special Reference to Plasma Proteins, Vol 1. Elsevier, Amsterdam-London-New York, pp 173–235

    Google Scholar 

  52. Jakubke H-D, Jeschkeit H (eds) (1981) Lexikon Biochemie, 2nd ed. Verlag Chemie, Weinheim

    Google Scholar 

  53. Leach BS, Collawn JF Jr, Fish WW (1980) Biochemistry 19:5741–5747

    Article  CAS  Google Scholar 

  54. Kempner ES, Miller JH, McCreery MJ (1986) Anal Biochem 156:140–146

    Article  CAS  Google Scholar 

  55. Peters T Jr (1985) Adv Prot Chem 37:161–245

    Article  CAS  Google Scholar 

  56. Trimble RB, Maley F (1977) J Biol Chem 252:4409–4412

    CAS  Google Scholar 

  57. Malamud D, Drysdale JW (1978) Anal Biochem 86:620–647

    Article  CAS  Google Scholar 

  58. Treffry A, Harrison PM (1979) Biochem J 181:709–716

    CAS  Google Scholar 

  59. Fischbach FA, Anderegg JW (1965) J Mol Biol 14:458–473

    CAS  Google Scholar 

  60. Crichton RR, Charloteaux-Wauters M (1987) Eur J Biochem 164:485–506

    Article  CAS  Google Scholar 

  61. Svensson B, Pedersen TG, Svendsen I, Sakai T, Ottesen M (1982) Carlsberg Res Commun 47:55–69

    CAS  Google Scholar 

  62. Clarke AJ, Svensson B (1984) Carlsberg Res Commun 49:559–566

    CAS  Google Scholar 

  63. Kronman MJ, Andreotti RE (1964) Biochemistry 3:1145–1151

    Article  CAS  Google Scholar 

  64. Andrews P (1965) Biochem J 96:595–606

    CAS  Google Scholar 

  65. Bloomfield VA (1983) Biopolymers 22:2141–2154

    Article  CAS  Google Scholar 

  66. Teng T-L, Harpst JA, Lee JC, Zinn A, CArlson DM (1976) Arch Biochem Biophys 176:71–81

    Article  CAS  Google Scholar 

  67. Christl P (1990) Diplomarbeit. University of Regensburg, FRG

    Google Scholar 

  68. Neumann NP, Lampen JO (1967) Biochemistry 6:468–475

    Article  CAS  Google Scholar 

Download references

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Authors and Affiliations

  1. Institut für Biophysik und Physikalische Biochemie, Universität Regensburg, Universitätsstraße 31, D-8400, Regensburg, FRG

    H. Durchschlag, R. Christl & R. Jaenicke

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  1. H. Durchschlag
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  2. R. Christl
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  3. R. Jaenicke
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W. Borchard

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© 1991 Dr. Dietrich Steinkopff Verlag GmbH & Co. KG

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Durchschlag, H., Christl, R., Jaenicke, R. (1991). Comparative determination of the particle weight of glycoproteins by SDS-PAGE and analytical ultracentrifugation. In: Borchard, W. (eds) Progress in Analytical Ultracentrifugation. Progress in Colloid & Polymer Science, vol 86. Steinkopff. https://doi.org/10.1007/BFb0115006

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  • DOI: https://doi.org/10.1007/BFb0115006

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  • Publisher Name: Steinkopff

  • Print ISBN: 978-3-7985-0910-8

  • Online ISBN: 978-3-7985-1683-0

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