Surface induced dissociation yields substructure of Methanosarcina thermophila 20S proteasome complexes

摘要

Native mass spectrometry (MS) and surface induced dissociation (SID) have been applied to study the stoichiometry and quaternary structure of non-covalent protein complexes. In this study, Methanosarcina thermophila 20S proteasome, which consists of four stacked heptameric rings (alpha(7)beta(7)beta(7)alpha(7) symmetry), has been selected to explore the SID dissociation pattern of a complicated stacked ring protein complex. SID produces both alpha and beta subunits while collision induced dissociation (CID) produces only highly charged alpha subunit. In addition, the charge reduced 20S proteasome produces the alpha(7)beta(7) fragment, reflecting the stacked ring topology of the complex. The combination of SID and charge reduction is shown to be a powerful tool for the study of protein complex structure. (C) 2014 Published by Elsevier B.V.