PT - JOURNAL ARTICLE AU - Boer Xie AU - Amika Sood AU - Robert J. Woods AU - Joshua S. Sharp TI - Quantitative Protein Topography Measurements by High Resolution Hydroxyl Radical Protein Footprinting Enable Accurate Molecular Model Selection AID - 10.1101/136929 DP - 2017 Jan 01 TA - bioRxiv PG - 136929 4099 - http://biorxiv.org/content/early/2017/05/11/136929.short 4100 - http://biorxiv.org/content/early/2017/05/11/136929.full AB - We report an integrated workflow that allows mass spectrometry-based high-resolution hydroxyl radical protein footprinting (HR-HRPF) measurements to accurately measure the absolute average solvent accessible surface area (<SASA>) of amino acid side chains. This approach is based on application of multi-point HR-HRPF, electron-transfer dissociation (ETD) tandem MS (MS/MS) acquisition, measurement of effective radical doses by radical dosimetry, and proper normalization of the inherent reactivity of the amino acids. The accuracy of the resulting <SASA> measurements was tested by using well-characterized protein models. Moreover, we demonstrated the ability to use <SASA> measurements from HR-HRPF to differentiate molecular models of high accuracy (< 3Å backbone RMSD) from models of lower accuracy (> 4Å backbone RMSD). The ability of <SASA> data from HR-HRPF to differentiate molecular model quality was found to be comparable to that of <SASA> data obtained from X-ray crystal structures, indicating the accuracy and utility of HR-HRPF for evaluating the accuracy of computational models.