Covalent binding of RNA with peptides in the structure of a specific complex using N-oxybenzotriazole oligoribonucleotide esters
Taran, E.A.; Ivanovskaia, M.G.; Gait, M.J.; Shabarova, Z.A.
Molekuliarnaia Biologiia 32(5): 832-839
1998
ISSN/ISBN: 0026-8984 PMID: 9914970 Document Number: 491653
A method was proposed for covalent RNA binding with peptides in a specific complex. The method is based on the use of oligoribonucleotide N-hydroxybenzotriazole esters. These RNA derivatives are able to phosphorylate protein amino groups in an aqueous medium. Synthetic RNA duplex containing protein binding site Tat of type I HIV was used as an example to show that the introduction of N-hydroxybenzotriazole at the RNA terminal phosphate group did not prevent the formation of a specific complex with peptide. Covalent binding of the activated RNA duplex with peptide takes place without additional external activation with the effectiveness of 20-30 %. In the absence of the specific complex, covalent binding does not take place when RNA duplex is replaced by a RNA-DNA hybrid or single-stranded RNA. Phosphamide nature of covalent bond between RNA and peptide was established that indicated the participation of Lys epsilon-amino group or peptide terminal alpha-amino group in the formation of a bond with RNA.