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

Warning

This publication doesn't include Faculty of Arts. It includes Central European Institute of Technology. Official publication website can be found on muni.cz.

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).
Related projects:	Centrum nových přístupů k bioanalýze a molekulární diagnostice