Methods for the large-scale synthesis of psoralen furan-side monoadducts and diadducts.
نویسندگان
چکیده
We report methods for the preparation of multimicromole quantities of extremely pure uniquely photo-adducted psoralen-DNA furan-side monoadducts and diadducts (cross-links). The methods use high-intensity krypton and argon ion lasers in the photoreactions and HPLC methods to purify the required oligonucleotides containing the photoadducts. With these methods we have synthesized 2-3 mumol of 8-mer psoralen furan-side monoadduct and diadduct. These methods allow one to generate large amounts of psoralenated DNA oligonucleotides and facilitate their study by NMR and x-ray crystallography.
منابع مشابه
Recent advances in the synthesis and structure determination of site specifically psoralen-modified DNA oligonucleotides.
We have developed novel methods for the preparation of multimicromole quantities of extremely pure, uniquely photoadducted psoralen-DNA cross-links, furan-side monoadducted DNA and pyrone-side monoadducts. Psoralen cross-linked and furan-side monoadducted DNA were produced by employing high intensity argon ion and krypton ion lasers as light sources. Pyrone-side monoadducts were prepared by bas...
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We have studied the wavelength dependence for the photoreversal of a monoadducted psoralen derivative, HMT [4'-(hydroxymethyl)-4,5',8-trimethylpsoralen], in a single-stranded deoxyoligonucleotide (5'-GAAGCTACGAGC-3'). The psoralen was covalently attached to the thymidine residue in the oligonucleotide as either a furan-side monoadduct, which is formed through the cycloaddition between the 4',5'...
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We have studied the photochemical reactions of 8-methoxypsoralen (&MOP) with calf thymus DNA. Analysis of the photoproducts formed was carried out by enzymatic digestion of the 8-MOP-modified DNA, followed by HPLC separation of photoadducts by high-pressure liquid chromatography (HPLC). The 4’,5’ (furan-side) monoadduct of &MOP bound to thymidine is converted to cross-liilked thymidine8-MOP-thy...
متن کاملPsoralens as photoactive probes of nucleic acid structure and function: organic chemistry, photochemistry, and biochemistry.
PERSPECTIVES AND UMMARY .............................................................. 1152 INTRODUCTION ..................................................................................... 1155 ORGANIC AND STRUCTURAL CHEMISTRY OF PSORALEN AND ITS ADDUCTS ................................................................... 1158 Naturally Occurring Psoralens ........................................
متن کاملRepair of 4,5',8-trimethylpsoralen monoadducts and cross-links by the Escherichia coli UvrABC endonuclease.
Using an oligonucleotide model substrate, we observed two unusual mechanisms of UvrABC endonuclease in the repair of 4,5',8-trimethylpsoralen monoadducts and crosslinks. (i) UvrABC endonuclease usually incises a psoralen monoadduct only on the damaged strand. However, for one of the monoadducts we studied, incision on the complementary undamaged strand was also observed at a very low frequency,...
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ورودعنوان ژورنال:
- Proceedings of the National Academy of Sciences of the United States of America
دوره 89 10 شماره
صفحات -
تاریخ انتشار 1992