Photochemical cross-linking of protein and DNA in chromatin. Synthesis and application of a photosensitive cleavable derivative of 9-aminoacridine with two photoprobes connected through a disulphide-containing linker

Research output: Contribution to journalJournal articleResearchpeer-review

A novel cleavable photo-cross-linking reagent, N-(2-methoxy-6-azidoacridin-9-yl)-N'-(4-azidobenzoyl)cystamine, for analysis of protein-nucleic acid interactions, has been synthesized. The reagent contains two photosensitive groups that can be activated sequentially. The azidoacridinyl moiety is sensitive to u.v. and visible light (lambda less than or equal to 450 nm), whereas the azidobenzoyl part needs higher-energy light (lambda less than or equal to 350 nm). Furthermore, the disulphide bridge connecting the two photoactive groups can be cleaved by reduction with mercaptans. The reagent is shown to induce cleavable cross-links between all five major histones and DNA in chromatin from Ehrlich ascites cells on activation with long-wavelength u.v. light (lambda greater than 300 nm) at an efficiency of approximately 3% of the added reagent.

Original languageEnglish
JournalBiochemical Journal
Volume223
Issue number2
Pages (from-to)519-26
Number of pages8
ISSN0264-6021
DOIs
Publication statusPublished - 15 Oct 1984

    Research areas

  • Azides/chemical synthesis, Chromatin/analysis, DNA, Electrophoresis, Polyacrylamide Gel, Histones, Photochemistry, Spectrophotometry, Ultraviolet Rays

ID: 203631879