Dataset Information

Fast Protein Footprinting by X-ray Mediated Radical Trifluoromethylation.

ABSTRACT: Synchrotron radiolysis generates hydroxyl radicals (^•OH) that are successful footprinting reagents. Here, we describe a new reagent for the synchrotron platform, the trifluoromethyl radical (^•CF₃). The radical is produced by ^•OH displacement of ^•CF₃ from sodium triflinate (Langlois reagent). Upon X-ray beam exposure, the reagent labels proteins extensively without any additional chemicals on a millisecond or shorter time scale. The ^•CF₃ is comparably reactive to ^•OH and produces footprinting information that complements that of ^•OH alone. This reagent in combination with ^•OH should enable novel chemistry for protein footprinting on the synchrotron platform.

SUBMITTER: Cheng M

PROVIDER: S-EPMC7486011 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Fast Protein Footprinting by X-ray Mediated Radical Trifluoromethylation.

Cheng Ming M Asuru Awuri A Kiselar Janna J Mathai George G Chance Mark R MR Gross Michael L ML

Journal of the American Society for Mass Spectrometry 20200421 5

Synchrotron radiolysis generates hydroxyl radicals (•OH) that are successful footprinting reagents. Here, we describe a new reagent for the synchrotron platform, the trifluoromethyl radical (•CF3). The radical is produced by •OH displacement of •CF3 from sodium triflinate (Langlois reagent). Upon X-ray beam exposure, the reagent labels proteins extensively without any additional chemicals on a millisecond or shorter time scale. Th ...[more]

PMID: 32255631

Similar Datasets

Project description:Measurements from hydroxyl radical footprinting (HRF) provide rich information about the solvent accessibility of amino acid side chains of a protein. Traditional HRF data analyses focus on comparing the difference in the modification/footprinting rate of a specific site to infer structural changes across two protein states, e.g., between a free and ligand-bound state. However, the rate information itself is not fully used for the purpose of comparing different protein sites within a protein on an absolute scale. To provide such a cross-site comparison, we present a new, to our knowledge, data analysis algorithm to convert the measured footprinting rate constant to a protection factor (PF) by taking into account the known intrinsic reactivity of amino acid side chain. To examine the extent to which PFs can be used for structural interpretation, this PF analysis is applied to three model systems where radiolytic footprinting data are reported in the literature. By visualizing structures colored with the PF values for individual peptides, a rational view of the structural features of various protein sites regarding their solvent accessibility is revealed, where high-PF regions are buried and low-PF regions are more exposed to the solvent. Furthermore, a detailed analysis correlating solvent accessibility and local structural contacts for gelsolin shows a statistically significant agreement between PF values and various structure measures, demonstrating that the PFs derived from this PF analysis readily explain fundamental HRF rate measurements. We also tested this PF analysis on alternative, chemical-based HRF data, showing improved correlations of structural properties of a model protein barstar compared to examining HRF rate data alone. Together, this PF analysis not only permits a novel, to our knowledge, approach of mapping protein structures by using footprinting data, but also elevates the use of HRF measurements from a qualitative, cross-state comparison to a quantitative, cross-site assessment of protein structures in the context of individual conformational states of interest.

Dataset Information

Fast Protein Footprinting by X-ray Mediated Radical Trifluoromethylation.

Publications

Fast Protein Footprinting by X-ray Mediated Radical Trifluoromethylation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets