A new mass spectrometer for structural proteomics and protein imaging

Proteins are large molecules associated with a vast array of chemical modifications, and which form loosely-bound complexes with themselves, other proteins and other molecules. Proteins work together as machines to perform all the processes required for life. Their functions are intrinsically linked to their overall structure. Mass spectrometry is an analytical technique which enables us to track proteins to understand more about how they work. The global analysis of protein structure is known as structural proteomics, and makes use of mass spectrometry.

The aim of this proposal is to install the most advanced mass spectrometer for structural proteomics at the University of Birmingham. The instrument offers high sensitivity, high mass accuracy, and high mass resolving power. The mass spectrometer will also be equipped with the latest ion mobility spectrometry technology, the FAIMS Pro. This device improves the sensitivity and separation capabilities, meaning that it is possible to delve deeper and wider into the proteome. Importantly, the instrument enables cross-linking experiments, providing new capabilities for mapping the architecture of protein complexes. Additionally, the instrument will bring new opportunities for improved mapping of intact protein distribution (mass spectrometry imaging) in tissue and in living microbial colonies.

The proposal brings together a broad range of researchers from the University of Birmingham and the wider Midlands (University of Nottingham, University of Leicester) from diverse disciplines to address several key strategic priorities for BBSRC (Healthy ageing across the life course, Combatting antimicrobial resistance, Sustainably enhancing agricultural production, Technology for the biosciences, and Data driven biology).

The aim of the proposal is to purchase an Orbitrap Fusion Lumos mass spectrometer. The instrument has the brightest ion source, a quadrupole filter with improved selectivity and ion transmission, higher capacity ETD fragmentation, and is equipped with advanced beam ion guide technology that reduces noise and improves robustness compared with previous instrument designs. The mass analyser provides resolution in excess of 500,000 FWHM and scan rates of 30 Hz meaning that this instrumentation is the best instrument for coupling with liquid chromatography; technology that is essential for structural proteomics experiments using top-down, middle-down and bottom-up protein identification. Additionally, the instrument will be equipped with FAIMS Pro technology, a cylindrical FAIMS device capable of separating gas-phase ions at atmospheric pressure. This device will improve sensitivity and separation capabilities of the mass spectrometer. This will be the first instrument of its kind in the UK, and one of only four worldwide. The PI has extensive experience of FAIMS, and therefore the University of Birmingham has the expertise essential to make the proposed research a success.

The University of Birmingham has a highly successful Advanced Mass Spectrometry Facility wherein over 100 research groups have benefited from its instrumentation (internal, national, international and industry). The enhanced features of the Orbitrap Fusion Lumos mass spectrometer will enable us to perform experiments that have previously been impossible, enabling us to break boundaries in biological research areas that involve direct protein detection from tissue, localisation of labile protein post-translational modifications, and mapping protein complex architecture by cross-linking mass spectrometry.

Who will benefit from the research?

The research proposal aims to develop and exploit the Orbitrap Fusion Lumos mass spectrometer with FAIMS Pro across a broad range of disciplines within the biological sciences. The pharmaceutical industry, the NHS and its patients, and analytical instrument manufacturers will benefit from this work.

How will they benefit from the research?

The proposed research has the potential to contribute significantly to the nation's health and wealth:

1) Pharmaceutical industry: The pharmaceutical industry is underpinned by analytical science. According to the 2016 report on 'Strength & Opportunity in the UK Life Sciences', commissioned by the Office for Life Sciences, the Life Sciences industry employs >200, 000 people in nearly 6000 companies, with an annual turnover of £61 bn. The biopharmaceutical industry employs >100,000 people in 2000 companies generating an annual turnover of £40 bn. The proposed equipment will support the research of Cooper who works closely with collaborators at AstraZeneca and UCB Pharma to develop new tools for drug discovery. Many of the projects to which the instrumentation will be applied relate to the drive to produce new antibiotics, either by identifying/understanding potential targets or developing new approaches for antibiotic synthesis. In the longer term, these projects will feed into the pharmaceutical industry.

2) NHS and its patients: The proposed instrumentation will be applied to understanding the virulence of Mycobacterium tuberculosis. Tuberculosis (TB) remains a major global health problem, reflected by its inclusion in the WHO's Millennium Development Goals ("reverse the spread of [tuberculosis] by 2015"). The 2011 case of Birmingham teenager Alina Sarag, whose TB infection was not diagnosed in time and ultimately caused her death, is a stark reminder that despite available antibiotics TB remains a serious threat in the UK.

The proposed instrument will also be applied to the understanding of oral health over the lifecourse. Oral health is a key indicator of overall health, well-being and quality of life. The World Health Organisation (WHO) prioritises research that targets both the young and elderly, since the oral health in these individuals is heavily linked to problems with diet and nutrition.

3) Analytical instrumentation manufacturers: The proposed research will involve close working with the instrument manufacturers to enable the technology to be developed and expanded further into new biological areas. The success of which will increase sales and thus be beneficial to the economy.