Releasing Nonperipheral Subunits from Protein Complexes in the Gas Phase.

Guanbo Wang, Lingxiao Chaihu, Meng Tian, Xinyang Shao, Rongrong Dai, Rob N de Jong, Deniz Ugurlar, Piet Gros, Albert J R Heck

Analytical chemistry 2020 Dec 15

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The quaternary structure is an important feature regulating protein function. Native mass spectrometry contributes to untangling quaternary structures by preserving the integrity of protein complexes in the gas phase. Tandem mass spectrometry by collision-induced dissociation (CID) can then be used to release subunits from these intact complexes, thereby providing structural information on the stoichiometry and topology. Cumulatively, such studies have revealed the preferred release of peripheral subunits during CID. In contrast, here we describe and focus on dissociation pathways that release nonperipheral subunits from hetero-complexes in CID at high collision energies. We find that nonperipheral subunits are ejected with a high propensity, as a consequence of sequential dissociation events, upon initial removal of peripheral subunits. Alternatively, nonperipheral subunits can be released directly from a charge-reduced or an elongated intact complex. As demonstrated here for a range of protein assemblies, releasing nonperipheral subunits under controlled conditions may provide unique structural information on the stoichiometry and topology of protein complexes.

Citation

Guanbo Wang, Lingxiao Chaihu, Meng Tian, Xinyang Shao, Rongrong Dai, Rob N de Jong, Deniz Ugurlar, Piet Gros, Albert J R Heck. Releasing Nonperipheral Subunits from Protein Complexes in the Gas Phase. Analytical chemistry. 2020 Dec 15;92(24):15799-15805