Cysteine-Specific Cu2+ Chelating Tags Used as Paramagnetic Probes in Double Electron Electron Resonance
| Autor: | Timothy F. Cunningham, Sunil Saxena, Christopher P. Jaroniec, Rajith J. Arachchige, Min Gao, Miriam R. Putterman, Matthew D. Shannon, Ishita Sengupta |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Nitroxide mediated radical polymerization
Magnetic Resonance Spectroscopy Analytical chemistry Cyclams Protein Structure Secondary Article chemistry.chemical_compound Protein structure Cyclen Heterocyclic Compounds Cations Materials Chemistry Side chain Cysteine Physical and Theoretical Chemistry Spin label Protein secondary structure Edetic Acid Chelating Agents biology Chemistry Proteins Nuclear magnetic resonance spectroscopy Surfaces Coatings and Films Crystallography biology.protein Spin Labels Protein G Copper |
| Zdroj: | The Journal of Physical Chemistry B. 119:2839-2843 |
| ISSN: | 1520-5207 1520-6106 |
| DOI: | 10.1021/jp5103143 |
| Popis: | Double electron electron resonance (DEER) is an attractive technique that is utilized for gaining insight into protein structure and dynamics via nanometer-scale distance measurements. The most commonly used paramagnetic tag in these measurements is a nitroxide spin label, R1. Here, we present the application of two types of high-affinity Cu(2+) chelating tags, based on the EDTA and cyclen metal-binding motifs as alternative X-band DEER probes, using the B1 immunoglobulin-binding domain of protein G (GB1) as a model system. Both types of tags have been incorporated into a variety of protein secondary structure environments and exhibit high spectral sensitivity. In particular, the cyclen-based tag displays distance distributions with comparable distribution widths and most probable distances within 1-3 Å when compared to homologous R1 distributions. The results display the viability of the cyclen tag as an alternative to the R1 side chain for X-band DEER distance measurements in proteins. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|