Double Nitroxide Labeling by Copper-Catalyzed Azide-Alkyne Cycloadditions with Noncanonical Amino Acids for Electron Paramagnetic Resonance Spectroscopy.
| Autor: | Widder P; Department of Chemistry and Konstanz Research School Chemical Biology , University of Konstanz , Konstanz , Germany., Berner F; Department of Chemistry and Konstanz Research School Chemical Biology , University of Konstanz , Konstanz , Germany., Summerer D; Faculty of Chemistry and Chemical Biology , TU Dortmund University , Dortmund , Germany., Drescher M; Department of Chemistry and Konstanz Research School Chemical Biology , University of Konstanz , Konstanz , Germany. |
|---|---|
| Jazyk: | angličtina |
| Zdroj: | ACS chemical biology [ACS Chem Biol] 2019 May 17; Vol. 14 (5), pp. 839-844. Date of Electronic Publication: 2019 May 01. |
| DOI: | 10.1021/acschembio.8b01111 |
| Abstrakt: | Electron paramagnetic resonance spectroscopy in combination with site-directed spin labeling (SDSL) is an important tool to obtain long-range distance restraints for protein structural research. We here study a variety of azide- and alkyne-bearing noncanonical amino acids (ncAA) in terms of protein single- and double-incorporation efficiency via nonsense suppression, metabolic stability, yields of nitroxide labeling via copper-catalyzed [3 + 2] azide-alkyne cycloadditions (CuAAC), and spectroscopic properties in continuous-wave and double electron-electron resonance measurements. We identify para-ethynyl-l-phenylalanine and para-propargyloxy-l-phenylalanine as suitable ncAA for CuAAC-based SDSL that will complement current SDSL approaches, particularly in cases in which essential cysteines of a target protein prevent the use of sulfhydryl-reactive spin labels. |
| Databáze: | MEDLINE |
| Externí odkaz: |
|