Photoreactivating Enzyme from Escherichia coli: Appearance of New Absorption on Binding to Ultraviolet Irradiated DNA

K. L. Wun, Alice Gih, John Clark Sutherland

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The photoreactivating enzyme, PRE, monomerizes pyrimidine dimers in DNA in a light requiring reaction (λ > 300 nm). However, the purified PRE from E. coli has no well-defined absorption band for λ > 300 nm. Using absorption difference spectroscopy, we show that when PRE is mixed with ultraviolet-irradiated DNA, new absorption appears in the spectral region required for catalysis. There is a concomitant decrease in the absorption of the mixture for wavelengths less than 300 nm. The hyperchromicity for λ > 300 nm is true absorption, not an artifact due to light scattering. Both the hyperchromicity (λ > 300 nm) and hypochromicity (λ < 300 nm) can be reversed by irradiation at 365 nm with identical first-order kinetics. We estimate the molar extinction coefficient of the new absorption to be 6900 ± 1400 at 350 nm. We conclude that the PRE from E. coli does not possess a distinct “chromophore” which by itself is entirely responsible for the absorption of photoreactivating light. Instead, new absorption results when PRE binds its substrate, dimer-containing DNA.

Original languageEnglish (US)
Pages (from-to)921-924
Number of pages4
JournalBiochemistry
Volume16
Issue number5
DOIs
StatePublished - Mar 1 1977
Externally publishedYes

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Deoxyribodipyrimidine Photo-Lyase
Escherichia coli
DNA
Pyrimidine Dimers
Chromophores
Dimers
Light scattering
Catalysis
Absorption spectra
Light
Irradiation
Spectroscopy
Wavelength
Kinetics
Substrates
Artifacts
Spectrum Analysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Photoreactivating Enzyme from Escherichia coli : Appearance of New Absorption on Binding to Ultraviolet Irradiated DNA. / Wun, K. L.; Gih, Alice; Sutherland, John Clark.

In: Biochemistry, Vol. 16, No. 5, 01.03.1977, p. 921-924.

Research output: Contribution to journalArticle

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