Site-specific protein immobilization by Staudinger ligation.
The Staudinger ligation between an azido-protein and a phosphinothioester-derivatized surface is demonstrated to be an effective means for the site-specific, covalent immobilization of a protein. Immobilization yields of >50% are obtained in <1 min, and immobilized proteins have >80% of their expected activity. No other method enables more rapid immobilization or a higher yield of active protein. Because azido-peptides and azido-proteins are readily attainable by synthesis, biosynthesis, or semisynthesis, the Staudinger ligation could be of unsurpassed utility in creating microarrays of functional peptides and proteins.
D000595:Amino Acid Sequence; D001386:Azides / Q000138:chemical synthesis / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D004800:Enzymes, Immobilized / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry*
D001386:Azides; D004800:Enzymes, Immobilized; D010446:Peptide Fragments; C023560:ribonuclease S-peptide; D011092:Polyethylene Glycols; D012260:Ribonucleases