New tools for structural biology: Native ambient mass spectrometry

Secondary Supervisor(s): Dr Aneika Leney

University of Registration: University of Birmingham

BBSRC Research Themes: Understanding the Rules of Life (Structural Biology)

Project Outline

Native ambient mass spectrometry (NAMS) is an emerging field that enables label-free characterisation of proteins and their complexes directly from their physiological environment. NAMS has the potential to be an enormously powerful tool for structural biology. For example, NAMS has the potential to provide structural information on a protein-drug complex, or a pathological protein aggregate such as found in Alzheimer’s Disease, directly from the tissue itself, providing unprecedented insight into fundamental biological processes. What’s more, NAMS provides this information in a spatially-defined manner. That is, NAMS may be applied for mass spectrometry imaging. The benefits of NAMS can be illustrated by considering a thin tissue section through a drug-treated tumour. In principle, NAMS would inform on drug-target interactions (i.e., is the drug binding to the correct protein?)) and location of drug target interactions (i.e., is the protein-drug complex in the tumour or healthy tissue?).

The Cooper lab is at the forefront of NAMS research and have demonstrated that intact protein complexes can be extracted from tissue sections by two complementary techniques: liquid extraction surface analysis (LESA) [1, 2] and nanospray desorption electrospray ionisation (nano-DESI) [3, 4]. Despite these promising results, the underlying mechanisms of extraction of native proteins remain unclear, as do the optimum sampling and analysis conditions. The aim of the project is to address these questions and to further develop these techniques for NAMS of a range of tissue types. The research will also involve integration of ion mobility spectrometry techniques with the NAMS techniques to further improve performance. Computational tools for data analysis (protein identification and visualisation of mass spectrometry imaging datasets) will be developed.

References

1. Hale, O.J., et al., J. Am. Soc. Mass Spectrom., 2020, 31, 873-879.

2. Griffiths, R.L., et al., Int. J. Mass Spectrom., 2017, 437, 23-29.

3. Hale, O.J., et al., Native Ambient Mass Spectrometry of an Intact Membrane Protein Assembly and Soluble Protein Assemblies Directly from Lens Tissue. 2022: University of Birmingham eData Repository.

4. Sisley, E.K., et al., J. Am. Chem. Soc., 2022, 144, 2120-2128.

In situ interrogation of small and novel large molecule drugs by native ambient mass spectrometry imaging

Secondary Supervisor(s): Dr Oliver Hale

University of Registration: University of Birmingham

BBSRC Research Themes:

• Integrated Understanding of Health (Pharmaceuticals)
• Understanding the Rules of Life (Structural Biology)

Project Outline

Native ambient mass spectrometry (NAMS) is an emerging field that enables label-free characterization of proteins and their complexes directly from their biological environment. NAMS has the potential to be an enormously powerful tool for drug discovery. For example, NAMS has the potential to provide structural information on a protein-drug complex directly from the tissue itself, providing unprecedented insight into fundamental biological processes. As NAMS provides this information in a spatially-defined manner, it can be applied for mass spectrometry imaging.

The Cooper lab is at the forefront of NAMS research and have demonstrated that intact protein-drug complexes can be extracted from tissue sections by two complementary techniques: liquid extraction surface analysis (LESA) and nanospray desorption electrospray ionisation (nano-DESI).

The aim of the project is to further develop and apply NAMS for in situ analysis of protein-drug interactions. Drug modalities to be investigated include novel drug modalities including large and small molecule drugs.

The project is in partnership with AstraZeneca, a long-term collaborator of the Cooper group.

Techniques

Mass spectrometry Liquid extraction surface analysis (LESA) Nanospray desorption electrospray ionisation (Nano-DESI), MALDI mass spectrometry imaging, Ion mobility spectrometry (FAIMS, TWIMS), Immunohistochemistry, Bioinformatics tools, visualisation software tools.