title: Chloroacetamide-Linked Nucleotides and DNA for Cross-Linking with Peptides and Proteins.
abstract: 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).
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Gilead Sciences & IOCB Research Center , Flemingovo namesti 2, 166 10 Prague 6, Czech Republic.
authors.forename: Agata
authors.initials: A
authors.lastname: Olszewska
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Gilead Sciences & IOCB Research Center , Flemingovo namesti 2, 166 10 Prague 6, Czech Republic.
authors.forename: Radek
authors.initials: R
authors.lastname: Pohl
authors.affiliation: Institute of Biophysics, Czech Academy of Sciences , Kralovopolska 135, 612 65 Brno, Czech Republic.
authors.forename: Marie
authors.initials: M
authors.lastname: Brázdová
authors.affiliation: Institute of Biophysics, Czech Academy of Sciences , Kralovopolska 135, 612 65 Brno, Czech Republic.|Central European Institute of Technology, Masaryk University , Kamenice 753/5, CZ-625 00 Brno, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Fojta
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Gilead Sciences & IOCB Research Center , Flemingovo namesti 2, 166 10 Prague 6, Czech Republic.|Department of Organic Chemistry, Faculty of Science, Charles University in Prague , Hlavova 8, 12843 Prague 2, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Hocek
mesh_terms: D000081:Acetamides / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004579:Electron Transport; D006639:Histidine / Q000737:chemistry; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
chemical_list: D000081:Acetamides; D009711:Nucleotides; D010455:Peptides; D011506:Proteins; C013874:chloroacetamide; D006639:Histidine; D004247:DNA; D003545:Cysteine