Developing a nanopore tweezer toolkit for analyzing kinase structural dynamics and screening allosteric inhibitors

The University of Massachusetts received a $396,854 Project Grant from the National Institute of General Medical Sciences (NIGMS) under the Biomedical Research and Research Training program (CFDA 93.859), effective September 1, 2025, through August 31, 2029. The award supports the development of a nanopore tweezer toolkit designed to analyze kinase structural dynamics and screen allosteric inhibitors. This research leverages protein nanopore sensor technology to enable real-time tracking of kinase movements triggered by ligand or inhibitor binding at 100-microsecond resolution, sustained for tens of minutes at the single-molecule level. The project will generate comprehensive energetic maps of conformational states in disease-associated kinases during substrate binding, catalytic activity, allosteric regulation, and inhibition.

The deliverables from this award include novel nanopore-based analytical tools and methodologies applicable to single-molecule enzymology, drug discovery, and personalized precision medicine. By advancing protein analysis capabilities through innovative nanopore approaches, this research aims to improve disease diagnostics and therapeutics while establishing new opportunities for fundamental structural biology research. The work is performed at the University of Massachusetts' facilities in Massachusetts.

Abstract

Protein nanopores are powerful sensors that enable a new class of single-molecule measurements, with applications ranging from fundamental science to commercial DNA sequencing. Our focus is on developing and applying novel nanopore approaches to tackle challenges in protein analysis, ultimately improving disease diagnostics and therapeutics. In this project, we are creating innovative nanopore tweezers capable of real-time tracking kinase movements triggered by ligand or inhibitor binding at a 100-microsecond resolution for tens of minutes at the single-molecule level. This work will generate comprehensive energetic maps of the conformational states of disease-associated kinases during substrate binding, catalytic activity, allosteric regulation, and inhibition. Overall, this study will introduce new tools and opportunities for studying structural dynamics, single-molecule enzymology, drug discovery, and personalized precision medicine.