Work

Separation and Collision Cross Section Measurements of Protein Complexes Afforded by a Modular Drift Tube Coupled to an Orbitrap Mass Spectrometer

Year: 2022

Type: article

Abstract: New developments in analytical technologies and biophysical methods have advanced the characterization of increasingly complex biomolecular assemblies using native mass spectrometry (MS). Ion mobility methods, in particular, have enabled a new dimension of structural information and analysis of proteins, allowing separation of conformations and providing size and shape insights based on collision cross sections (CCSs). Based on the concepts of absorption-mode Fourier transform (aFT) multiplexing ion mobility spectrometry (IMS), here, a modular drift tube design proves capable of separating native-like proteins up to 148 kDa with resolution up to 45. Coupled with high-resolution Orbitrap MS, binding of small ligands and cofactors can be resolved in the mass domain and correlated to changes in structural heterogeneity observed in the ion-neutral CCS distributions. We also demonstrate the ability to rapidly determine accurate CCSs for proteins with 1-min aFT-IMS-MS sweeps without the need for calibrants or correction factors.

Source: Analytical Chemistry

DOI: 10.1021/acs.analchem.2c01653

Authors: Sarah N. Sipe, James D. Sanders, Tobias Reinecke, Brian H. Clowers, Jennifer S. Brodbelt

Institutions: The University of Texas at Austin, Washington State University

Language: English

Topic: Mass Spectrometry Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funders: National Science Foundation, Welch Foundation, National Institute of General Medical Sciences

Awards: Ultraviolet Photodissociation Mass Spectrometry, G4226750041, Innovative Native Ion Mobility Approaches for Transformational Measurements in Structural Biology