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Related Experiment Videos

DNA photoreactivating enzyme from human tissues.

S E Ogut1, S M D'Ambrosio, M Samuel

  • 1Biology Department, Brookhaven National Laboratory, Upton, NY 11973.

Journal of Photochemistry and Photobiology. B, Biology
|October 1, 1989
PubMed
Summary

Human tissues contain photoreactivating enzyme (PRE) activity that repairs DNA damage using light. This enzyme activity was found in fetal skin, kidney, lung, liver, brain, and intestine, as well as neonatal foreskin.

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Area of Science:

  • Molecular biology
  • Biochemistry
  • Genetics

Background:

  • DNA damage is a constant threat to cellular integrity.
  • Photoreactivating enzymes (PREs) are known to repair DNA damage using visible light.
  • Understanding PRE distribution and characteristics in human tissues is crucial for DNA repair research.

Purpose of the Study:

  • To quantitate photoreactivating enzyme activity in various human fetal tissues and neonatal foreskin.
  • To characterize the properties of these enzyme activities, including molecular size, heat stability, and protease sensitivity.
  • To confirm if these activities represent true photoreactivating enzymes.

Main Methods:

  • Quantitation of enzyme activity in homogenates of human fetal skin, kidney, lung, liver, brain, intestine, and neonatal foreskin.

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  • Assay of DNA dimer photolysis using wavelengths greater than 320 nm.
  • Enzyme inactivation studies using trypsin, pronase, and heat (65°C).
  • Determination of molecular size by filtration.

    • Main Results:

    • Photoreactivating enzyme activity was detected in all examined human tissues.
    • Two distinct molecular size activities were observed: >10,000 Da and <10,000 Da.
    • The enzyme activities were heat-labile and sensitive to protease digestion.
    • Catalytic photolysis of pyrimidine dimers was confirmed, independent of tryptophan.

    Conclusions:

    • Human fetal and neonatal tissues possess photoreactivating enzyme (PRE) activity.
    • The characterized properties align with those of a true photoreactivating enzyme, indicating its presence and function in human tissues.
    • These findings contribute to the understanding of DNA repair mechanisms in humans.