Open AccessDetermination of solvent content in cavities in IL-1β using experimentally phased electron density
Author(s) -
M.L. Quillin,
Paul T. Wingfield,
Brian W. Matthews
Publication year - 2006
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0609442104
Subject(s) - solvent , chemistry , electron density , molecule , crystallography , polar , phase (matter) , spectroscopy , chemical physics , analytical chemistry (journal) , electron , physics , organic chemistry , quantum mechanics , astronomy
The extent to which water is present within apolar cavities in proteins remains unclear. In the case of interleukin-1β (IL-1β), four independent structures solved by x-ray crystallography indicate that water is not present in the central apolar cavity. In contrast, results from NMR spectroscopy suggest that water has high occupancy within the cavity but is positionally disordered, making it undetectable by standard crystallographic methods. A theoretically based crystallographic-phase refinement technique also suggested that there was the equivalent of two fully occupied water molecules within the apolar cavity. To resolve these discrepancies we sought to obtain an experimentally phased electron density map that was free of possible bias caused by mathematical modeling of the protein or the solvent. By combining native diffraction data with multiple wavelength anomalous data from a platinum derivative, accurate phases were obtained. Using these experimental phases, we estimate that occupancy of the apolar cavity in IL-1β by solvent is close or equal to zero. Polar cavities in the protein that contain ordered solvent molecules serve as internal controls.