ABSTRACT
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.
Subject(s)
Azides/chemistry , Escherichia coli Proteins/chemistry , Escherichia coli/chemistry , Organophosphorus Compounds/chemistry , Peptide Elongation Factors/chemistry , Peptides/chemistry , Amino Acids/chemistry , Nuclear Magnetic Resonance, Biomolecular , Staining and LabelingABSTRACT
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.
Subject(s)
Phenanthrolines/chemistry , Proteins/chemistry , Ruthenium/chemistry , Molecular Structure , Proteins/analysisABSTRACT
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.
