Determination of protein-protein interactions by matrix-assisted laser desorption/ionization mass spectrometry.
A number of different procedures have been developed for use with matrix-assisted laser desorption/ionization mass spectrometry (MALDI/MS) for the analysis of non-covalent protein-protein complexes. These include use of specific matrix and laser combinations, accumulation of "first shot" spectra, modification of pH and solvent conditions during sample preparation and use of cross-linking agents to attach the monomers covalently to each other in the complex. The results have shown the techniques to be effective with some but not all complexes, although cross-linking is the most successful. The physical and chemical nature of the complex is critical and therefore a diversity of approaches is recommended for such studies.
Farmer
T B
TB
Department of Biochemistry and Mass Spectrometry Research Center, Vanderbilt Unitversity School of Medicine, Nashville, Tennessee 37232-6400, USA.
Caprioli
R M
RM
D000818:Animals; D002417:Cattle; D003432:Cross-Linking Reagents; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D013048:Specimen Handling / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D003432:Cross-Linking Reagents; D011506:Proteins