Metastable ion formation and disparate charge separation in the gas-phase dissection of protein assemblies studied by orthogonal time-of-flight mass spectrometry.
The dissection of specific and nonspecific protein complexes in the gas phase is studied by collisionally activated decomposition. In particular, the gas phase dissection of multiple protonated homodimeric Human Galectin I, E. Coli Glyoxalase I, horse heart cytochrome c, and Hen egg Lysozyme have been investigated. Both the Human Galectin I and E. Coli Glyoxalase I enzymes are biologically active as a dimer, exhibiting molecular weights of approximately 30 kDa. Cytochrome c and Lysozyme are monomers, but may aggregate to some extent at high protein concentrations. The gas phase dissociation of these multiple protonated dimer assemblies does lead to the formation of monomers. The charge distribution over the two concomitant monomers following the dissociation of these multiple protonated dimers is found to be highly dissimilar. There is no evident correlation between the solution phase stability of the dimeric proteins and their gas-phase dissociation pattern. Additionally, in the collisionally activated decomposition spectra diffuse ion signals are observed, which are attributed to monomer ions formed via slow decay of the collisionally activated dimer ions inside the reflectron time-of-flight. Although, the formation of these diffuse metastable ions may complicate the interpretation of collisionally activated decomposition mass spectra, especially when studying noncovalent protein complexes, a simple mathematical equation may be used to reveal their origin and pathway of formation.
Versluis
C
C
Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, The Netherlands.
van der Staaij
A
A
Stokvis
E
E
Heck
A J
AJ
de Craene
B
B
D000465:Algorithms; D003574:Cytochrome c Group / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry; D037161:Galectins; D006388:Hemagglutinins / Q000737:chemistry*; D006801:Humans; D007791:Lactoylglutathione Lyase / Q000737:chemistry*; D013058:Mass Spectrometry; D009113:Muramidase / Q000737:chemistry*
D003574:Cytochrome c Group; D037161:Galectins; D006388:Hemagglutinins; D009113:Muramidase; D007791:Lactoylglutathione Lyase