Nucleotides, 2'-deoxyribonucleoside triphosphates (dNTPs), and DNA probes bearing reactive chloroacetamido group linked to nucleobase (cytosine or 7-deazadaenine) through a propargyl tether were prepared and tested in cross-linking with cysteine- or histidine-containing peptides and proteins. The chloroacetamide-modifed dNTPs proved to be good substrates for DNA polymerases in the enzymatic synthesis of modified DNA probes. Modified nucleotides and DNA reacted efficiently with cysteine and cysteine-containing peptides, whereas the reaction with histidine was sluggish and low yielding. The modified DNA efficiently cross-linked with p53 protein through alkylation of cysteine and showed potential for cross-linking with histidine (in C277H mutant of p53).

Central European Institute of Technology Masaryk University Kamenice 753 5 CZ 625 00 Brno Czech Republic

Department of Organic Chemistry Faculty of Science Charles University Prague Hlavova 8 12843 Prague 2 Czech Republic

Institute of Biophysics Czech Academy of Sciences Kralovopolska 135 612 65 Brno Czech Republic

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences Gilead Sciences and IOCB Research Center Flemingovo namesti 2 166 10 Prague 6 Czech Republic

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