Site-specific protein modifications through pyrroline-carboxy-lysine residues
| Autor: | Jan Grünewald, Qian Fan, David H. Jones, Ansgar Brock, Linda Okach, Scott A. Lesley, Tiffany Crossgrove, Lisa Quinn, Weijia Ou, Hsien-Po Chiu, Tetsuo Uno, Susan E. Cellitti, Bernhard H. Geierstanger, Paula Patterson, Xueshi Hao |
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| Rok vydání: | 2011 |
| Předmět: |
Multidisciplinary
Oligonucleotide Lysine technology industry and agriculture Pyrrolysine Proteins macromolecular substances Biological Sciences equipment and supplies musculoskeletal system medicine.disease_cause Small molecule Culture Media chemistry.chemical_compound Biotin chemistry Biochemistry Transfer RNA Escherichia coli medicine Pyrroles Nuclear Magnetic Resonance Biomolecular Ethylene glycol |
| Zdroj: | Proceedings of the National Academy of Sciences. 108:10437-10442 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1105197108 |
| Popis: | Pyrroline-carboxy-lysine (Pcl) is a demethylated form of pyrrolysine that is generated by the pyrrolysine biosynthetic enzymes when the growth media is supplemented with D-ornithine . Pcl is readily incorporated by the unmodified pyrrolysyl-tRNA/tRNA synthetase pair into proteins expressed in Escherichia coli and in mammalian cells. Here, we describe a broadly applicable conjugation chemistry that is specific for Pcl and orthogonal to all other reactive groups on proteins. The reaction of Pcl with 2-amino-benzaldehyde or 2-amino-acetophenone reagents proceeds to near completion at neutral pH with high efficiency. We illustrate the versatility of the chemistry by conjugating Pcl proteins with poly(ethylene glycol)s, peptides, oligosaccharides, oligonucleotides, fluorescence, and biotin labels and other small molecules. Because Pcl is genetically encoded by TAG codons, this conjugation chemistry enables enhancements of the pharmacology and functionality of proteins through site-specific conjugation. |
| Databáze: | OpenAIRE |
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