Open AccessClass II DNA photolyase from Arabidopsis thaliana contains FAD as a cofactor
Author(s) -
Kleiner Oliver,
Butenandt Jens,
Carell Thomas,
Batschauer Alfred
Publication year - 1999
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.1999.00590.x
Subject(s) - photolyase , pyrimidine dimer , arabidopsis , arabidopsis thaliana , chromophore , cofactor , dna , biochemistry , escherichia coli , enzyme , biology , chemistry , dna repair , photochemistry , gene , mutant
The major UV‐B photoproduct in DNA is the cyclobutane pyrimidine dimer (CPD). CPD‐photolyases repair this DNA damage by a light‐driven electron transfer. The chromophores of the class II CPD‐photolyase from Arabidopsis thaliana , which was cloned recently [Taylor, R., Tobin, A. & Bray, C. (1996) Plant Physiol . 112 , 862; Ahmad, M., Jarillo, J.A., Klimczak, L.J., Landry, L.G., Peng, T., Last, R.L. & Cashmore, A.R. (1997) Plant Cell 9 , 199–207], have not been characterized so far. Here we report on the overexpression of the Arabidopsis CPD photolyase in Escherichia coli as a 6 × His‐tag fusion protein, its purification and the analysis of the chromophore composition and enzymatic activity. Like class I photolyase, the Arabidopsis enzyme contains FAD but a second chromophore was not detectable. Despite the lack of a second chromophore the purified enzyme has photoreactivating activity.