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

0464736 - ÚOCHB 2017 RIV US eng J - Journal Article
Olszewska, Agata - Pohl, Radek - Brázdová, Marie - Fojta, Miroslav - Hocek, Michal
Chloroacetamide-Linked Nucleotides and DNA for Cross-Linking with Peptides and Proteins.
Bioconjugate Chemistry. Roč. 27, č. 9 (2016), s. 2089-2094. ISSN 1043-1802. E-ISSN 1520-4812
R&D Projects: GA ČR GBP206/12/G151
Institutional support: RVO:61388963 ; RVO:68081707
Keywords : tumor suppressor p53 * Diels-Alder reaction * reductive amination
OECD category: Biochemistry and molecular biology; Organic chemistry (BFU-R)
Impact factor: 4.818, year: 2016
Method of publishing: Open access
http://pubs.acs.org/doi/full/10.1021/acs.bioconjchem.6b00342

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).
Permanent Link: http://hdl.handle.net/11104/0263512