This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Protein-ligand interactions by mass spectrometry, titration, and H/D exchange (PLIMSTEX) is a mass spectrometric method for detg. assocn. consts. and binding stoichiometry for interactions of proteins with various ligands, as well as for quantifying the conformational changes assocd. with ligand binding to proteins. The assocn. consts. detd. with PLIMSTEX agree with literature values within a factor of six, establishing its validity for protein interactions involving metal ions, small org. mols., peptides, and proteins. PLIMSTEX provides soln., not gas-phase, properties by taking advantage of ESI and MALDI mass spectrometry to measure accurately the mass of a protein as it undergoes amide H/D exchange. The approach sidesteps the problem of relating gas-phase abundances of the protein or protein-ligand complex ions to their soln. concns. With on-column concn. and desalting, high picomole quantities of proteins are sufficient for reproducible mass detection, and the concn. of the protein can be as low as 10-8 M. It is amenable to different protein/ligand systems in physiol. relevant media. No specially labeled protein or ligand is needed. PLIMSTEX offers minimal perturbation of the binding equil. because it uses no denaturants, no addnl. spectroscopy or reaction probes, and no phys. sepn. of ligand and protein during binding. In this research, we are developing, testing, and extending further the method.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
2P41RR000954-33
Application #
8168666
Study Section
Special Emphasis Panel (ZRG1-BCMB-K (40))
Project Start
2010-03-10
Project End
2010-12-31
Budget Start
2010-03-10
Budget End
2010-12-31
Support Year
33
Fiscal Year
2010
Total Cost
$56,325
Indirect Cost
Name
Washington University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
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