Open AccessChloroacetamide-Linked Nucleotides and DNA for Cross-Linking with Peptides and Proteins
Author(s) -
Agata Olszewska,
Radek Pohl,
Marie Brázdová,
Miroslav Fojta,
Michal Hocek
Publication year - 2016
Publication title -
bioconjugate chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.279
H-Index - 172
eISSN - 1520-4812
pISSN - 1043-1802
DOI - 10.1021/acs.bioconjchem.6b00342
Subject(s) - chemistry , cysteine , nucleotide , nucleobase , dna , histidine , biochemistry , cytosine , deoxyribonucleoside , dna polymerase , enzyme , gene
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).