Cysteine-specific Cu2+ chelating tags used as paramagnetic probes in double electron electron resonance.
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| Abstract | 
   :  
              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.  | 
        
| Year of Publication | 
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              2015 
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| Journal | 
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              The journal of physical chemistry. B 
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| Volume | 
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              119 
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| Issue | 
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              7 
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| Number of Pages | 
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              2839-43 
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| Date Published | 
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              2015 
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| ISSN Number | 
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              1520-6106 
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| URL | 
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              https://dx.doi.org/10.1021/jp5103143 
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| DOI | 
   :  
              10.1021/jp5103143 
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| Short Title | 
   :  
              J Phys Chem B 
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