Title: Next-Generation Protein Stability Measurements in the Absence of Bulk Solvent
Abstract:
Medicines of the future will rely heavily upon our ability to quickly assess the structures and stabilities of complex macromolecular machines and evaluate the influence of large libraries of conformationally-selective small molecule binders. In addition, the ever-growing and complex array of protein and nucleic acid-based biotherapeutics similarly demands improved tools that can rapidly detect changes in therapeutic structure upon target binding, alterations in formulation, or stress. Current structural biology tools lack the throughput and sensitivity to meet these challenges. In this presentation, I will discuss recent developments surrounding collision induced unfolding (CIU) methods that aim to bridge this technology gap. CIU uses ion mobility-mass spectrometry (IM-MS) to measure the stability and unfolding pathways of gas-phase proteins, without the need for covalent labels or tagging, and consumes 10-100 times less sample than almost any other label-free technology. Recent developments in high-throughput CIU screening methods, their ability to track small alterations in structure over a wide array of biotherapeutics, and software developments that seek to enhance CIU information content, will be discussed.
Bio:
Brandon T. Ruotolo is currently Professor and Associate Chair for Research in the Department of Chemistry, University of Michigan. Ruotolo is also the faculty director of the UM Biological Mass Spectrometry Facility and a faculty affiliate of both the Department of Biological Chemistry and the Computational Medicine and Bioinformatics Program within the UM Medical School. He earned his B.S. in Chemistry from Saint Louis University in 1999, and his Ph.D. from Texas A&M University in 2004 under the direction of David H. Russell. He did his post-doctoral work at the University of Cambridge with Dame Carol V. Robinson and was awarded the inaugural Waters Research Fellowship in 2008. Brandon moved to the University of Michigan in 2009, where he began his independent career. The Ruotolo research group at the University of Michigan seeks to enable breakthroughs in structural biology and drug discovery by leveraging the potential of ion mobility-mass spectrometry (IM-MS) for the comprehensive, 3D structural analysis of the proteome. To this end, Ruotolo and his team have studied the role of solvation on biomolecular structure, introduced collision induced unfolding (CIU) – a new fingerprinting technology capable of detecting the structural state of protein-ligand complexes and biotherapeutics, developed software packages for the enhanced interpretation and throughput of IM-MS and CIU data, as well as investigated the structural consequences of small molecule drug-like compounds on amyloid-related peptides and proteins. Ruotolo’s work has resulted in ca. 170 peer-reviewed publications and many awards, including the Research Award from the American Society for Mass Spectrometry (2011), the Eli Lilly Young Investigator Award (2013), the NSF CAREER award (2013), the Protein Science Young Investigator Award (2018), the Agilent Thought Leader Award (2018), the Biemann Medal from the American Society for Mass Spectrometry (2023), and the Ken Standing Award from the University of Manitoba and ETP Symposium Inc (2024).
Keywords: Mass spectrometry, Protein folding, Biotherapeutics, Pharmaceuticals, Structural Biology