title: High-resolution crystal structure of spin labelled (T21R1) azurin from Pseudomonas aeruginosa: a challenging structural benchmark for in silico spin labelling algorithms.
abstract: BACKGROUND
  EPR-based distance measurements between spin labels in proteins have become a valuable tool in structural biology. The direct translation of the experimental distances into structural information is however often impaired by the intrinsic flexibility of the spin labelled side chains. Different algorithms exist that predict the approximate conformation of the spin label either by using pre-computed rotamer libraries of the labelled side chain (rotamer approach) or by simply determining its accessible volume (accessible volume approach). Surprisingly, comparisons with many experimental distances have shown that both approaches deliver the same distance prediction accuracy of about 3 Å.
  
  
  RESULTS
  Here, instead of comparing predicted and experimental distances, we test the ability of both approaches to predict the actual conformations of spin labels found in a new high-resolution crystal structure of spin labelled azurin (T21R1). Inside the crystal, the label is found in two very different environments which serve as a challenging test for the in silico approaches.
  
  
  CONCLUSIONS
  Our results illustrate why simple and more sophisticated programs lead to the same prediciton error. Thus, a more precise treatment of the complete environment of the label and also its interactions with the environment will be needed to increase the accuracy of in silico spin labelling algorithms.
authors.forename: Nicole
authors.initials: N
authors.lastname: Florin
authors.forename: Olav
authors.initials: O
authors.lastname: Schiemann
authors.affiliation: Institute for Physical and Theoretical Chemistry, University of Bonn, Wegelerstr, 12, Bonn, NRW 53115, Germany. hagelueken@pc.uni-bonn.de.
authors.forename: Gregor
authors.initials: G
authors.lastname: Hagelueken
mesh_terms: D000465:Algorithms; D001400:Azurin / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D019295:Computational Biology; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D013113:Spin Labels
chemical_list: D001426:Bacterial Proteins; D013113:Spin Labels; D001400:Azurin