Electrospray ionization of native membrane proteins proceeds via a charge equilibration step.

ABSTRACT: Electrospray ionization mass spectrometry is increasingly applied to study the structures and interactions of membrane protein complexes. However, the charging mechanism is complicated by the presence of detergent micelles during ionization. Here, we show that the final charge of membrane proteins can be predicted by their molecular weight when released from the non-charge reducing saccharide detergents. Our data indicate that PEG detergents lower the charge depending on the number of detergent molecules in the surrounding micelle, whereas fos-choline detergents may additionally participate in ion-ion reactions after desolvation. The supercharging reagent sulfolane, on the other hand, has no discernible effect on the charge of detergent-free membrane proteins. Taking our observations into

SUBMITTER: Yen HY 

PROVIDER: S-EPMC8972943 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature