With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, and with partial co-funding from the Division of Molecular and Cellular Biosciences, Professor Brandon Ruotolo and his group at the University of Michigan are developing powerful new tools for characterizing the 3-dimensional structure of proteins - key molecules in life processes. Specifically, the Ruotolo group is expanding the capabilities of "collision induced unfolding" (CIU), an approach which studies protein structures by quickly removing the water which surrounds protein molecules in their normal environment, then induces their unfolding while monitoring their size and shape using a state-of-the-art tool known as ion mobility-mass spectrometry (IM-MS). The approach has the potential to revolutionize the detail and acquisition speed with which protein stability can be evaluated. Potential applications include the rapid analysis of biotherapeutic proteins and their mode of operation. The work provides excellent training for students interested in research at the interface between chemical measurements and the pharmaceutical sciences.
The research addresses three main goals: (1) creating a complete physical description of factors impacting the peak width for traveling-wave-type ion mobility separators, and validating the resulting predictions against standards; (2) developing next-generation software that incorporates the interpretation of IM peak width into CIU analysis workflows in the form of highly-parameterized multi-modal Gaussian fitting in both two and three dimensions; and (3) creating parallelized, multi-dimensional CIU methods capable of rapidly tracking the unfolding patterns of protein mixtures across many charge states simultaneously, even in the presence of significant chemical noise. The new CIU tools are being tested using therapeutic proteins of significance to the biotechnology sector.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
