RESUMO
Chemoselective modification of biomolecules: The reaction between 2-cyanoethyl phosphoramidites and azides is economical and can be performed in different solvents, including aqueous buffers. The course of the reaction with azido-modified amino acids, peptides or proteins and different label molecules was followed by (31)P NMR spectroscopy.
Assuntos
Azidas/química , Proteínas de Escherichia coli/química , Escherichia coli/química , Compostos Organofosforados/química , Fatores de Alongamento de Peptídeos/química , Peptídeos/química , Aminoácidos/química , Ressonância Magnética Nuclear Biomolecular , Coloração e RotulagemRESUMO
The bioorthogonal and chemoselective fluorescence labelling of several cell-free synthesized proteins containing a site-specifically incorporated azido amino acid was possible using different alkyne-functionalized Ru(II) bathophenanthroline complexes. We were able to achieve a selective labelling even in complex mixtures of proteins despite the fact that ruthenium dyes normally show a high tendency for unspecific interactions with proteins and are commonly used for total staining of proteins. Since the employed Ru complexes are extremely robust, photo-stable and highly sensitive, the approach should be applicable to the production of labelled proteins for single molecule spectroscopy and fluorescence-based interaction studies.
Assuntos
Fenantrolinas/química , Proteínas/química , Rutênio/química , Estrutura Molecular , Proteínas/análiseRESUMO
A new method for the oxidation of nucleoside phosphite triester into phosphate triester under nonbasic and nonaqueous conditions using NBS-DMSO in CH(3)CN has been developed. The utility of this method for solution- and solid-phase synthesis of oligonucleotide is demonstrated.
