Formation and characterization of protein-protein complexes in vacuo.
Gas-phase reactions between multiply charged positive and negative protein ions are carried out in a quadrupole ion trap mass spectrometer. The ions react with one another by proton transfer and complex formation. Proton transfer products and complexes are formed via competitive processes in single ion/ion encounters. The relative contributions of proton transfer versus complex formation are dependent upon the charges of the ions as well as other characteristics of the ions yet to be clearly delineated. No fragmentation of covalent bonds of the protein reactants is observed. A model that considers the trajectories associated with ion/ion interactions appears to hold the most promise in accounting for the results. The formation of bound ion/ion orbits appears to play an important role in determining overall reaction kinetics as well as the distribution of ion/ion reaction products. Tandem mass spectrometry is used to compare protein complexes formed in the gas-phase with those formed initially in solution and subsequently liberated by electrospray; it is shown that both forms of complex dissociate similarly, but the complexes formed in the gas phase can retain a "memory" of their method of formation.
Wells
J Mitchell
JM
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907-2084, USA.
Chrisman
Paul A
PA
McLuckey
Scott A
SA
D000818:Animals; D002417:Cattle; D003574:Cytochrome c Group / Q000737:chemistry*; D019281:Dimerization; D006736:Horses; D007477:Ions; D007700:Kinetics; D011522:Protons; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D025801:Ubiquitin / Q000737:chemistry*; D014618:Vacuum; D014835:Volatilization
D003574:Cytochrome c Group; D007477:Ions; D011522:Protons; D025801:Ubiquitin