Bankmann M, Brendel M
Institut für Mikrobiologie der J. W. Goethe-Universität, Frankfurt/Main, F.R.G.
J Photochem Photobiol B. 1989 Feb;3(1):33-52. doi: 10.1016/1011-1344(89)80019-3.
We present methods for the determination of UVA-induced binding of 8-methoxypsoralen (8-MOP) to nucleic acids and protein and for a quantitative assay of radioactively labelled 8-MOP plus UVA induced DNA photoproducts in the yeast Saccharomyces cerevisiae. For the dose range up to 60 kJ m-2, with a wild-type survival of 1% or higher, binding to DNA is 100-fold and to RNA 10- to 20-fold more efficient than that to protein. Between 20% and 65% of the 8-MOP binds to macromolecules that are neither nucleic acids nor protein. The number of DNA-bound 8-MOP molecules for the haploid genome rises from 14 (unirradiated control) to 349 at the highest UVA exposure dose (60 kJ m-2). Gel chromatography reveals three types of DNA thymine photoproduct, the pyrone-side monoadducts, the furan-side monoadducts and the diadducts. Among these, pyrone-side monoadducts always constitute the smallest fraction, regardless of whether the treatment is with in vitro or in vivo 8-MOP plus UVA.
我们介绍了用于测定8-甲氧基补骨脂素（8-MOP）与核酸及蛋白质的紫外线A（UVA）诱导结合的方法，以及用于定量检测酿酒酵母中放射性标记的8-MOP加UVA诱导的DNA光产物的方法。在高达60 kJ m-2的剂量范围内，野生型存活率为1%或更高时，8-MOP与DNA的结合效率比与蛋白质的结合效率高100倍，与RNA的结合效率比与蛋白质的结合效率高10至20倍。8-MOP中有20%至65%与既不是核酸也不是蛋白质的大分子结合。单倍体基因组中与DNA结合的8-MOP分子数量从14个（未辐照对照）增加到最高UVA暴露剂量（60 kJ m-2）时的349个。凝胶色谱法揭示了三种类型的DNA胸腺嘧啶光产物，即吡喃侧单加合物、呋喃侧单加合物和双加合物。在这些光产物中，无论处理是采用体外还是体内的8-MOP加UVA，吡喃侧单加合物始终占最小比例。
