Native Mass Spectrometry Guided Structural Biology Center

We propose conversion of the P41 Resource for Native Mass Spectrometry Guided Structural Biology (2018- ) to an RM1 Biomedical Technology Development and Dissemination (BTDD) Center. The nMSSB Center will produce robust, universally-available, transformative technology for characterizing the assembly and disassembly of macromolecular protein complexes, including the definition of their compositions, PTMs, stoichiometry, structure/architecture/topology, flexibility/stability, and their conformational/binding-induced changes under different conditions. The inclusion of an integrated workflow for native mass spectrometry (nMS) will enable, complement, and expand what is possible with other structural biology (SB) tools. We propose three highly synergistic Technology Development Projects (TDPs) that build from our productive P41 Technology Research and Development. Feasibility for the TDPs has been established through preliminary data and publications in high-quality journals, as well as through the licensing of one version of our SID technology by an MS vendor (Waters Corp.), release of a desalting column (based on our online buffer exchange progress) by another vendor (Thermo) in 2022, and the funding of two SBIR grants (eMSion and Protein Metrics). We continue and expand our relationships with multiple MS vendors/ sustainability partners, biopharma collaborators/industrial advisory board members, and add two technology partners (Ivanov and Coon). Ten geographically and topically/scientifically diverse Driving Biomedical Projects (DBPs) have been selected to drive technology development and extend nMS to leading biomedical laboratories that are tackling e.g., cancer, viral infections, and neurological disorders. Our three TDPs focus on different vendor-neutral tools coupled in multiple ways to maximize future nMS options. TDP1 optimizes a panel of solution-phase separation and measurement tools, including the expansion of two-column online buffer exchange, capillary electrophoresis, and variable temperature electrospray ionization. TDP2 develops improved mass selection using digital quadrupole technology and improved resolution Fourier transform ion mobility (IM) spectrometry, couples surface induced dissociation with multiple IM and MS analyzers including charge detection mass spectrometry, and develops computational methods to use energy-resolved nMS data and IM to identify “near-native” computational models. TDP3 develops collision- and surface-induced unfolding to screen biomolecular stability, adapts TDP technologies for vendor-specific complex down/native-omics approaches, and constructs integrated computations that utilize synergistic MS restraints to predict protein quaternary structures. Our administration & management structure provides leadership, coordinates communication/collaboration, and drives sustainability. Our community engagement plan continues dissemination and training to include publications, technology transfer to beta testers and vendors, an annual nMS short course, virtual and hands-on workshops, and multiple “sustainability sites” where nMS workflows will be integrated with other SB tools for non-expert and expert users.