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.
This is a preview of subscription content, log in via an institution.
Preview
Unable to display preview. Download preview PDF.
Abbreviations
- AUC:
-
analytical ultracentrifugation
- HSSE:
-
high-speed sedimentation equilibrium
- SDS-PAGE:
-
sodium dodecyl sulfate polyacrylamide gel electrophoresis
- SEC:
-
size-exclusion chromatography
References
Hockwin O (ed) (1985) Biochemic des Auges. Ferdinand Enke Verlag, Stuttgart
Berman ER (1991) Biochemistry of the Eye, Plenum Press, New York
Summers L, Wistow G, Narebor M, Moss D, Lindley P, Slingsby C, Blundell T, Bartunik H, Bartels K (1984) Pept Protein Rev 3:147
Hockwin O (ed) (1976) Progress of Lens Biochemistry Research. Doc Ophthalmol Proc Ser, Vol 8. Dr W Junk by Publishers, The Hague
Lerman S (1980) Radiant Energy and the Eye. Macmillan, New York
Harding J (1991) Cataract: Biochemistry, Epidemiology and Pharmacology. Chapman & Hall, London
Durchschlag H, Fochler C, Feser B, Hausmann S, Seroneit T, Swientek M, Swoboda E, Winklmair A, Wlček C, Zipper P (1996) Radiat Phys Chem 47:501
von Sonntag C (1987) The Chemical Basis of Radiation Biology. Taylor & Francis, London
Kiefer J (ed) (1977) Ultraviolette Strahlen. Walter de Gruyter, Berlin
Fochler C (1996) Thesis, University of Regensburg
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 91–104
Yphantis DA (1964) Biochemistry 3:297
Durchschlag H, Durchschlag G (1977) Hoppe-Seyler's Z Physiol Chem 358:228
Sacket DL, Wolff J (1987) Anal Biochem 167:228
Durchschlag H, Christl P, Jaenicke R (1991) Progr Colloid Polym Sci 86:41
Durchschlag H, Jaenicke R (1983) Int J Biol Macromol 5:143
Habeeb AFSA (1972) Meth Enzymol 25:457
Thannhauser TW, Konishi Y, Scheraga HA (1987) Meth Enzymol 143:115
Borkman RF, McLaughlin J (1995) Photochem Photobiol 62:1046
Jaenicke R (1994) Naturwissenschaften 81:423
Jaenicke R (1996) FASEB J 10:84
Author information
Authors and Affiliations
Editor information
Rights and permissions
Copyright information
© 1997 Dr. Dietrich Steinkopff Verlag GmbH & Co. KG
About this paper
Cite this paper
Fochler, C., Durchschlag, H. (1997). Investigation of irradiated eye-lens proteins by analytical ultracentrifugation and other techniques. In: Jaenicke, R., Durchschlag, H. (eds) Analytical Ultracentrifugation IV. Progress in Colloid & Polymer Science, vol 107. Steinkopff. https://doi.org/10.1007/BFb0118020
Download citation
DOI: https://doi.org/10.1007/BFb0118020
Published:
Publisher Name: Steinkopff
Print ISBN: 978-3-7985-1106-4
Online ISBN: 978-3-7985-1656-4
eBook Packages: Springer Book Archive