Combining Surface-Induced Dissociation and Charge Detection Mass Spectrometry to Reveal the Native Topology of Heterogeneous Protein Complexes

22 May 2023, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Charge detection mass spectrometry (CDMS) enables the direct mass measurement of heterogenous samples on the megadalton scale, as the charge state for a single ion is determined simultaneously with mass to charge ratio (m/z). Surface-induced dissociation (SID) is an effective activation method to dissociate the noncovalent protein complexes without extensive gas-phase restructuring, producing various subcomplexes reflective of the native protein topology. Here, we demonstrate that using CDMS after SID on the Orbitrap platform offers subunit connectivity, topology, proteoform information, and relative interfacial strengths of the intact macromolecular assemblies. SID dissects the capsids (~3.7 MDa) of adeno-associated viruses (AAVs) into trimer-containing fragments (3mer, 6mer, 9mer, 15mer, etc) that can be detected by the individual ion mass spectrometry (I2MS) approach on Orbitrap instruments. SID coupled to CDMS provides unique structural insights into heterogenous assembles that are not obtained by traditional MS measurements.

Keywords

charge detection mass spectrometry
surface-induced dissociation
adeno-associated viruses
structural biology
CDMS
SID
AAV

Supplementary materials

Title
Description
Actions
Title
Supplementary information
Description
Supplementary information to the main manuscript titled: Combining Surface-Induced Dissociation and Charge Detection Mass Spectrometry to Reveal the Native Topology of Heterogeneous Protein Complexes
Actions

Comments

Comments are not moderated before they are posted, but they can be removed by the site moderators if they are found to be in contravention of our Commenting Policy [opens in a new tab] - please read this policy before you post. Comments should be used for scholarly discussion of the content in question. You can find more information about how to use the commenting feature here [opens in a new tab] .
This site is protected by reCAPTCHA and the Google Privacy Policy [opens in a new tab] and Terms of Service [opens in a new tab] apply.