Electrospray droplet exposure to polar vapors: delayed desolvation of protein complexes

Fragile non-covalent complexes are susceptible to dissociation upon introduction into and transmission through the mass spectrometer. The exposure of nanoelectrospray droplets to various polar vapors, which are introduced into the curtain gas, is shown to stabilize non-covalent protein complexes even under relatively energetic ion transfer conditions. This study probes the mechanism by which polar vapor exposure appears to stabilize non-covalent protein complex ions in the gas phase.

Holomyoglobin and hemoglobin were dissolved in either aqueous 1 mM ammonium acetate or ammonium bicarbonate solutions and ionized via nanoelectrospray ionization in the positive polarity. Polar vapors were entrained within the counter-current drying gas and exposed to nanoelectrospray droplets for circa 1 ms within the interface of a quadrupole/time-of-flight mass spectrometer. Mass spectra were acquired using various voltage gradients within the mass spectrometer.

In the absence of added reagent vapors, significant fragmentation of holomyoglobin ions is noted with high voltage gradients for ions either entering or departing q0, a transmission quadrupole closely coupled to the skimmer exit. However, upon the introduction of reagent vapors, essentially 100% of the holomyoglobin complex can be preserved. Significant stabilization is noted at both relatively high q0 entrance and exit gradients when ions are transmitted through q0. These results indicate that upon vapor exposure the holomyoglobin ions are not completely desolvated as they enter or exit q0 under normal ion transmission conditions.

The apparent stabilization of protein complexes and other non-covalent complexes noted here and elsewhere is attributed to the delayed desolvation of the ions. This allows the solvated ions to be transmitted through relatively high voltage gradients without disrupting the non-covalent interactions holding the complexes together.