Skip to main content
Log in

DNA-Binding proteins of psoriatic scales

I. Isolation by Affinity Chromatography and Characterisation by SDS Slab Gel Electrophoresis

  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

Summary

DNA binding proteins (DBP) are shown to be engaged in the regulatory functions of the genome. Since the accelerated epidermopoesis in psoriasis indicates changed gene activities, the examination of DBP possibly could help to clarify the pathological processes. Psoriatic scales were used to elaborate the method. DBP was isolated from a protein extract by affinity chromatography with DNA-cellulose according to Alberts et al. About 3-5% of postribosomal crude protein (PCP) extract had an affinity to DNA. The separation of DBP on SDS polyacrylamide slab gels showed a characteristic spectrum of proteins with molecular weights from 12.7 to 150 × 103 daltons. The protein band spectrum of the concentrated DBP fraction was different from that of the crude extract. These observations show the isolated DBP to represent an own group of water soluble proteins with DNA binding abilities.

Zusammenfassung

Bei der Aufklärung von Regulationsvorgängen am Genom wurden die chromosomalen DNA-affinen Proteine (DBP) als wesentliche Faktoren erkannt. Da die gesteigerte Epidermopoese bei der Psoriasis auf veränderte Genaktivitäten hinweist, könnten Untersuchungen der DBP ein Weg zur molekularbiologischen Klärung der pathologischen Vorgänge bei dieser Dermatose sein. Zur Ausarbeitung der Methodik dienten Psoriasisschuppen. Die Isolierung der DBP erfolgte aus einem Proteinextrakt affinitätschromatographisch mit DNA-Cellulose nach Alberts et al. Unter den Versuchsbedingungen besaßen 3-5% der Rohextraktproteine Affinität zur DNA. Die Auftrennung der DBP im SDS-Plattengel ergab ein charakteristisches Spektrum an Proteinen mit Molekulargewichten von 12700–150000. Das Bandenspektrum der angereicherten DBP-Fraktion unterschied sich hierbei von dem des Rohextraktes. Somit stellen die DBP eine eigene Gruppe an löslichen Proteinen mit erhaltener DNA-Affinität dar.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Alberts, B. M., Amadio, F. J., Jenkins, M., Gutmann, E. D., Ferris, F. L.: Studies with DNA-cellulose chromatography. I. DNA-binding proteins from E. coli. Cold Spring Harbour Symp. Quant. Biol.33, 289–304 (1968)

    Google Scholar 

  2. Burton, K.: Biochem. J.62, 315 (1956)

    Google Scholar 

  3. Chae, C.-B.: Reconstitution of chromatin-mode of reassociation of chromosomal proteins. Biochem.14, 900–906 (1975)

    Google Scholar 

  4. Dewair, M. A., Matthaei, H.: Tissue specificity of non-histone proteins from human chromatin. Molec. gen. Genet.146, 309–312 (1976)

    Google Scholar 

  5. Dunker, A. K., Rueckert, R. R.: Observations of molecular weight determinations on polyarylamid gel. J. Biol. Chem.244, 5074–5080 (1969)

    Google Scholar 

  6. Fellenberg, G.: Chromosomale proteine, p. 27–57. Stuttgart: E. Ulmer 1974

    Google Scholar 

  7. Francksen, H., Garadi, R.: Complete recovery of proteins in concentrators and some drawlochs, revealed by polyacrylamide gel electrophoresis. Z. Analyt. Chem.271, 340–344 (1974)

    Google Scholar 

  8. Jost, E., Lennox, R., Harris, H.: Affinity chromatography of DNA-binding proteins from human, murine and man-mouse hybrid cell lines. J. Cell Sci.18, 41–65 (1975)

    Google Scholar 

  9. Kostraba, N. C., Montagna, R. M., Wang, T. Y.: Study of the loosely bound non-histone chromatin proteins. J. Biol. Chem.250, 1548–1555 (1975)

    Google Scholar 

  10. Kostraba, N. C., Wang, T. Y.: Inhibition of transcription in vitro by a non-histone protein isolated from Ehrlich ascites tumor chromatin. J. Biol. Chem.250, 8938–8942 (1975)

    Google Scholar 

  11. Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Nature (Lond.)227, 680–685 (1970)

    Google Scholar 

  12. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Kolorimetrische Proteinbestimmung mit dem Folinschen Phenolreagenz. J. Biol. Chem.193, 265 (1951)

    Google Scholar 

  13. Mainwaring, W. I. P., Renni, P. S., Keekn, Y.: The androgenic regulation of prostate proteins with a high affinity for DNA. Biochem. J.156, 253–264 (1976)

    Google Scholar 

  14. Peter, G., Brand, A., Peter, R.: Studies on the species, amount and base composition of nucleic acids in psoriatic scales and normal horny layer. Dermatologica (Basel)151, 333–341 (1975)

    Google Scholar 

  15. Reimer, G., Ramb, G., Leonhardi, G.: Isolation and physico-chemical characterisation of the DNA from normal human skin and psoriatic scales. Arch. Derm. Res.256, 241–246 (1976)

    Google Scholar 

  16. Stein, G. H.: Differences in DNA-binding proteins isolated from normal and transformed human cells. Exp. Cell Res.99, 115–125 (1976)

    Google Scholar 

  17. Stein, G., Kleinsmith, L. J.: Chromosomal proteins and their role in the regulation of gene expression. New York: Academic Press 1976

    Google Scholar 

  18. Tashiro, L., Kurokowa, S.: Chromatin conformation as affected by shearing and by the removal of a portion of non-histone proteins. Febs Letters59, 250–253 (1975)

    Google Scholar 

  19. Teng, C. S., Teng, C. T., Allfrey, U. G.: Studies of nuclear proteins. J. Biol. Chem.246, 3597–3609 (1971)

    Google Scholar 

  20. Weber, K., Osborn, M.: The reliability of molecular weight determination by SDS-polyacrylamide gel electrophoresis. J. Biol. Chem.244, 4406–4412 (1969)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This work was supported by a grant of the Deutsche Forschungsgemeinschaft

We are greatly indebted to Mrs. M. Mentzer for her competent assistance

Rights and permissions

Reprints and permissions

About this article

Cite this article

Reimer, G., Leonhardi, G. & Schwulera, V. DNA-Binding proteins of psoriatic scales. Arch. Derm. Res. 260, 39–46 (1977). https://doi.org/10.1007/BF00558012

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00558012

Key words

Navigation