1. ABNOH-Linked Nucleotides and DNA for Bioconjugation and Cross-linking with Tryptophan-Containing Peptides and Proteins.
- Author
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Spampinato A, Leone DL, Pohl R, Tarábek J, Šoltysová M, Polák M, Kádek A, Sýkorová V, Řezáčová P, and Hocek M
- Subjects
- Proteins chemistry, Cross-Linking Reagents chemistry, DNA-Directed DNA Polymerase metabolism, DNA-Directed DNA Polymerase chemistry, Tryptophan chemistry, DNA chemistry, Peptides chemistry, Nucleotides chemistry
- Abstract
Reactive N-hydroxy-9-azabicyclo[3.3.1]nonane (ABNOH) linked 2'-deoxyuridine 5'-O-mono- and triphosphates were synthesized through a CuAAC reaction of ABNOH-PEG
4 -N3 with 5-ethynyl-dUMP or -dUTP. The modified triphosphate was used as substrate for enzymatic synthesis of modified DNA probes with KOD XL DNA polymerase. The keto-ABNO radical reacted with tryptophan (Trp) and Trp-containing peptides to form a stable tricyclic fused hexahydropyrrolo-indole conjugates. Similarly modified ABNOH-linked nucleotides reacted with Trp-containing peptides to form a stable conjugate in the presence but surprisingly even in the absence of NaNO2 (presumably through activation by O2 ). The reactive ABNOH-modified DNA probe was used for bioconjugations and crosslinking with Trp-containing peptides or proteins., (© 2024 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)- Published
- 2024
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