Genetically encoded dihydroxyphenylalanine coupled with tyrosinase for strain promoted labeling
| Autor: | Sundarapandian Ashokraj, Numbi Ramudu Kamini, P. Ganesan, Ganesh Shanmugam, A. George, Mohan Indhu, Niraikulam Ayyadurai |
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| Rok vydání: | 2021 |
| Předmět: |
Tyrosinase
Clinical Biochemistry Pharmaceutical Science Apoptosis Biochemistry law.invention law Drug Discovery Benzoquinones Moiety Humans Molecular Biology Cells Cultured Molecular Structure Chemistry Monophenol Monooxygenase Organic Chemistry Genetic code Recombinant Proteins Dihydroxyphenylalanine Recombinant DNA Click chemistry Molecular Medicine Click Chemistry Annexin A5 Genetic Engineering Macromolecule Fluorescent tag |
| Zdroj: | Bioorganicmedicinal chemistry. 50 |
| ISSN: | 1464-3391 |
| Popis: | Protein modifications through genetic code engineering have a remarkable impact on macromolecule engineering, protein translocation, protein-protein interaction, and cell biology. We used the newly developed molecular biology approach, genetic code engineering, for fine-tuning of proteins for biological availability. Here, we have introduced 3, 4-dihydroxy-l-phenylalanine in recombinant proteins by selective pressure incorporation method for protein-based cell labeling applications. The congener proteins treated with tyrosinase convert 3, 4-dihydroxy-l-phenylalanine to dopaquinone for strain-promoted click chemistry. Initially, the single-step Strain-Promoted Oxidation-Controlled Cyclooctyne-1,2-quinone Cycloaddition was studied using tyrosinase catalyzed congener protein and optimized the temporally controlled conjugation with (1R,8S,9s)-Bicyclo[6.1.0]non-4-yn-9-ylmethanol. Then, the feasibility of tyrosinase-treated congener annexin A5 with easily reactive quinone functional moiety was conjugated with fluorescent tag dibenzocyclooctyne-PEG4-TAMRA for labeling of apoptotic cells. Thus, the congener proteins-based products demonstrate selective cell labeling and apoptosis detection in EA.hy926 cells even after the protein modifications. Hence, genetic code engineering can be coupled with click chemistry to develop various protein-based fluorescent labels. |
| Databáze: | OpenAIRE |
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