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Parallel Chemoselective Profiling for Mapping Protein Structure.
- Source :
-
Cell chemical biology [Cell Chem Biol] 2020 Aug 20; Vol. 27 (8), pp. 1084-1096.e4. Date of Electronic Publication: 2020 Jul 09. - Publication Year :
- 2020
-
Abstract
- Solution-based structural techniques complement high-resolution structural data by providing insight into the oft-missed links between protein structure and dynamics. Here, we present Parallel Chemoselective Profiling, a solution-based structural method for characterizing protein structure and dynamics. Our method utilizes deep mutational scanning saturation mutagenesis data to install amino acid residues with specific chemistries at defined positions on the solvent-exposed surface of a protein. Differences in the extent of labeling of installed mutant residues are quantified using targeted mass spectrometry, reporting on each residue's local environment and structural dynamics. Using our method, we studied how conformation-selective, ATP-competitive inhibitors affect the local and global structure and dynamics of full-length Src kinase. Our results highlight how parallel chemoselective profiling can be used to study a dynamic multi-domain protein, and suggest that our method will be a useful addition to the relatively small toolkit of existing protein footprinting techniques.<br />Competing Interests: Declaration of Interests The authors declare no competing interests.<br /> (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Subjects :
- Adenosine Triphosphate chemistry
Adenosine Triphosphate metabolism
Binding, Competitive
Cysteine chemistry
HEK293 Cells
Humans
Molecular Dynamics Simulation
Mutagenesis, Site-Directed
PTEN Phosphohydrolase chemistry
PTEN Phosphohydrolase genetics
PTEN Phosphohydrolase metabolism
Protein Binding
Protein Kinase Inhibitors chemistry
Protein Kinase Inhibitors metabolism
Tandem Mass Spectrometry
Ubiquitin chemistry
Ubiquitin genetics
Ubiquitin metabolism
src-Family Kinases genetics
src-Family Kinases metabolism
Peptide Mapping methods
src-Family Kinases antagonists & inhibitors
Subjects
Details
- Language :
- English
- ISSN :
- 2451-9448
- Volume :
- 27
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Cell chemical biology
- Publication Type :
- Academic Journal
- Accession number :
- 32649906
- Full Text :
- https://doi.org/10.1016/j.chembiol.2020.06.014