Chloroacetamide-Linked Nucleotides and DNA for Cross-Linking with Peptides and Proteins

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Authors	OLSZEWSKA Agata POHL Radek BRAZDOVA Marie FOJTA Miroslav HOCEK Michal
Year of publication	2016
Type	Article in Periodical
Magazine / Source	BIOCONJUGATE CHEMISTRY
MU Faculty or unit	Central European Institute of Technology
Citation
Web	http://pubs.acs.org/doi/abs/10.1021/acs.bioconjchem.6b00342
Doi	http://dx.doi.org/10.1021/acs.bioconjchem.6b00342
Field	Biochemistry
Keywords	TUMOR-SUPPRESSOR P53; DIELS-ALDER REACTION; REDUCTIVE AMINATION; ENZYMATIC-SYNTHESIS; LYSINE RESIDUES; CLICK CHEMISTRY; BINDING; TRIPHOSPHATES; PROBES; AZIDE
Description	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).
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