Advertisement

Investigation of irradiated eye-lens proteins by analytical ultracentrifugation and other techniques

  • C. Fochler
  • H. Durchschlag
Biological Systems
Part of the Progress in Colloid & Polymer Science book series (PROGCOLLOID, volume 107)

Abstract

Crystallins, the major eye lens proteins, were investigated after preceding irradiation with X-rays or UV light by using analytical ultracentrifugation and other physicochemical and analytical techniques. Sedimentation velocity and sedimentation equilibrium runs of unirradiated and irradiated samples of the crude extract and individual fractions of calf-lens crystallins (α, β H, β L, γ) were performed under varying experimental conditions. While by the impact of radiation sedimentation coefficients remain essentially unchanged, high-speed sedimentation equilibrium experiments revealed significant changes of the molar masses of the respective crystallin species, depending on the crystallin class and the nature of the radiation used. The γ-crystallins turned out to be the most stable species against radiation-induced damages. UV absorption and fluorescence spectroscopy, size-exclusion chromatography, SDS electrophoresis, viscometry, and the analysis of SH and SS contents provided useful complementary information. In conclusion, several radiation damages could be registered: aggregation, crosslinking, dissociation, fragmentation, destruction of aromatic amino acids, cysteines and cystines. The OH scavenger formate and the chaperone α-crystallin turned out to protect the crystallin crude extract to some extent against X-ray-induced aggregation phenomena.

Key words

Eye-lens proteins crystallins radiation effects radioprotection analytical ultracentrifugation comparative studies 

Abbreviations

AUC

analytical ultracentrifugation

HSSE

high-speed sedimentation equilibrium

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEC

size-exclusion chromatography

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Hockwin O (ed) (1985) Biochemic des Auges. Ferdinand Enke Verlag, StuttgartGoogle Scholar
  2. 2.
    Berman ER (1991) Biochemistry of the Eye, Plenum Press, New YorkGoogle Scholar
  3. 3.
    Summers L, Wistow G, Narebor M, Moss D, Lindley P, Slingsby C, Blundell T, Bartunik H, Bartels K (1984) Pept Protein Rev 3:147Google Scholar
  4. 4.
    Hockwin O (ed) (1976) Progress of Lens Biochemistry Research. Doc Ophthalmol Proc Ser, Vol 8. Dr W Junk by Publishers, The HagueGoogle Scholar
  5. 5.
    Lerman S (1980) Radiant Energy and the Eye. Macmillan, New YorkGoogle Scholar
  6. 6.
    Harding J (1991) Cataract: Biochemistry, Epidemiology and Pharmacology. Chapman & Hall, LondonGoogle Scholar
  7. 7.
    Durchschlag H, Fochler C, Feser B, Hausmann S, Seroneit T, Swientek M, Swoboda E, Winklmair A, Wlček C, Zipper P (1996) Radiat Phys Chem 47:501CrossRefGoogle Scholar
  8. 8.
    von Sonntag C (1987) The Chemical Basis of Radiation Biology. Taylor & Francis, LondonGoogle Scholar
  9. 9.
    Kiefer J (ed) (1977) Ultraviolette Strahlen. Walter de Gruyter, BerlinGoogle Scholar
  10. 10.
    Fochler C (1996) Thesis, University of RegensburgGoogle Scholar
  11. 11.
    Bloemendal H, Zweers A (1976) In: Hockwin O (ed) Progress of Lens Biochemistry Research. Doc Ophthalmol Proc Ser, Vol 8. Dr W Junk by Publishers, The Hague, pp 91–104Google Scholar
  12. 12.
    Yphantis DA (1964) Biochemistry 3:297PubMedCrossRefGoogle Scholar
  13. 13.
    Durchschlag H, Durchschlag G (1977) Hoppe-Seyler's Z Physiol Chem 358:228Google Scholar
  14. 14.
    Sacket DL, Wolff J (1987) Anal Biochem 167:228CrossRefGoogle Scholar
  15. 15.
    Durchschlag H, Christl P, Jaenicke R (1991) Progr Colloid Polym Sci 86:41CrossRefGoogle Scholar
  16. 16.
    Durchschlag H, Jaenicke R (1983) Int J Biol Macromol 5:143CrossRefGoogle Scholar
  17. 17.
    Habeeb AFSA (1972) Meth Enzymol 25:457Google Scholar
  18. 18.
    Thannhauser TW, Konishi Y, Scheraga HA (1987) Meth Enzymol 143:115PubMedGoogle Scholar
  19. 19.
    Borkman RF, McLaughlin J (1995) Photochem Photobiol 62:1046PubMedCrossRefGoogle Scholar
  20. 20.
    Jaenicke R (1994) Naturwissenschaften 81:423PubMedGoogle Scholar
  21. 21.
    Jaenicke R (1996) FASEB J 10:84PubMedGoogle Scholar

Copyright information

© Dr. Dietrich Steinkopff Verlag GmbH & Co. KG 1997

Authors and Affiliations

  • C. Fochler
    • 1
  • H. Durchschlag
    • 2
  1. 1.GSFNational Research Center for Environment and HealthNeuherbergGermany
  2. 2.Institute of Biophysics and Physical BiochemistryUniversity of RegensburgRegensburgGermany

Personalised recommendations