... understanding life in molecular detail

Prof Alex Breeze

NMR, Structural and Chemical Biology, Protein-Protein Interactions, Cancer Drug Discovery

We combine state-of-the-art NMR spectroscopy with other structural and biophysical methods to interrogate both normal and disease biology at a molecular level. An important facet of this approach is the use of fragment-based methods to develop compounds that can be used as selective chemical biology probes. In some cases, these compounds may also form starting-points for developing therapeutic agents against important diseases such as cancer and bacterial or parasitic infections.

Current major projects include:
  • Fibroblast growth-factor receptor signalling and dynamics
  • Guanine-nucleotide exchange factors in disease and development
  • Ras pathway inhibition via novel allosteric mechanisms
  • Targeting Trypanosomal infections through unique PPIs

Our interests lie in the use of cutting-edge NMR approaches to interrogate structure, dynamics and interactions of biological macromolecules in normal and diease biology, with particular emphasis on cancer and bacterial or parasitic infections. Insights into the molecular-level mechanisms of disease biology can often be gleaned from studying the interactions of selective drugs or tool compounds with disease-associated proteins. We are combining techniques developed in many years’ experience of commercial drug discovery – for example, fragment-based inhibitor design using NMR screening – with structural and biophysical analysis of the way in which these compounds perturb the structure and dynamics of their target proteins.

We are interested in cell signalling through the fibroblast growth-factor receptor (FGFR) pathway and its role in development and cancer. In collaboration with Prof Matilda Katan (UCL) and Profs Adrian Goldman, Margaret Knowles and John Ladbury (Leeds), we are using NMR, EM and other techniques to understand how oncogenic and other activating mutations in the FGFR intracellular domain alter the normal regulation of FGFR activity. Our approach relies on the use of potent drug molecules that selectively stabilize different conformations of the protein as tools for understanding the effects of mutations on function. We are also collaborating with Prof Michele Vendruscolo (Cambridge) to combine experimental data from NMR with computational metadynamics simultations to understand the energetic drivers of conformational selection and allostery in kinases.

In normal cells, proteins of the Ras family of small GTPases rely on a complementary array of guanine nucleotide exchange factors (GEFs) to load them with the cofactor GTP, thus switching them to the ‘on’ state, whence they can signal to cells to grow and divide in response to external stimuli. In many cancers, this process is subverted by mutations in Ras that reduce or abolish reliance on GEFs and abrogate the normal mechanisms that replace GTP with GDP (the ‘off’ state of the Ras switch); this can result in permanently ‘on’ signals to cells to grow and divide – a hallmark of cancer. In addition, a number of developmental disorders including Noonan Syndrome are characterized by mutations in GEFs including the hSOS1 protein. In collaboration with Prof Mike Waring (Newcastle), we are using an array of structural and chemical biology techniques to investigate and understand how GEF interactions with normal and mutated Ras proteins and with other regulatory factors might be exploited for treating cancer and developmental diseases.

The adapter protein Grb2 and its close homologue Grb3-3 modulate SOS activity by binding to the C-terminal Pro-rich domain. In collaboration with Prof John Ladbury (Leeds) we are investigating the mechanism whereby Grb3-3 can dampen SOS activation by opposing Grb2-mediated recruitment. Since Grb3-3 expression levels are altered in certain cancers, understanding this mechanism may provide novel therapeutic opportunities.

In another avenue of investigation, we are interested in protein:protein interactions (PPIs) as potential drug targets. In collaboration with Prof John McCarthy (Warwick) and Prof Andy Wilson (Chemistry / Astbury, Leeds) we are attempting to understand how differences in the protein:protein interactions involved in translation initiation between higher eukaryotes and Trypanosomatids might be exploited for developing selective drugs against a debilitating class of developing-world diseases including Chagas disease; this project also involves collaborators in Brazil (Fiocruz Institute, Curitiba). A number of other PPI targets are also under investigation in the Breeze, Wilson, Tomlinson, Edwards and Nelson groups in the Astbury Centre.

BioNMR in Leeds benefits from state-of-the-art, cryoprobe-equipped instrumentation including a new 950 MHz and fully upgraded 750 and 600 MHz spectrometers, thanks to a >£17M investment in the Astbury BioStructure Laboratory from the University of Leeds and the Wellcome Trust. In particular, our 950 MHz is the first in the world to be equipped with a newly-developed 15N/13C direct detection-optimized cryoprobe, affording unique capabilities for studying large complexes and intrinsically disordered or highly mobile proteins.

Detailed research programme                  Close ▲

Professor of Biomolecular NMR
BSc (Imperial College) PhD (Imperial College)

SERC Postdoctoral Research Fellow (Oxford University) 1988-1992
Team Leader, Biomolecular NMR (Zeneca/AstraZeneca) 1991-2002
Associate Director, Biophysics (AstraZeneca) 2007-2010
Principal Scientist, Protein Structure and Biophysics (AstraZeneca) 20

Miall 10.28
School of Molecular and Cellular Biology
0113 343 0087

Selected Publications

  1. Patani H, Bunney TD, Thiyagarajan N, Norman RA, Ogg D, Breed J, Ashford P, Potterton A, Edwards M, Williams SV, Thomson GS, Pang CS, Knowles MA, Breeze AL, Orengo C, Phillips C & Katan M (2016) Landscape of activating cancer mutations in FGFR kinases and their differential responses to inhibitors in clinical use. Oncotarget Mar 16. doi: 10.18632/oncotarget.8132

  2. Klein T, Vajpai N, Phillips JJ, Davies G, Holdgate GA, Phillips C, Tucker JA, Norman RA, Scott AD, Higazi DR, Lowe D, Thompson GS & Breeze AL (2015) Structural and dynamic insights into the energetics of activation loop rearrangement in FGFR1 kinase. Nat. Commun. 6, 7877

  3. Vo U, Vajpai N, Flavell L, Bobby R, Breeze AL, Embrey KJ & Golovanov AP (2016) Monitoring Ras Interactions with the Nucleotide Exchange Factor Son of Sevenless (Sos) Using Site-specific NMR Reporter Signals and Intrinsic Fluorescence. J. Biol. Chem. 291, 1703-1718

  4. Ma J, McLeod S, MacCormack K, Sriram S, Gao N, Breeze AL & Hu J (2014) Real-time monitoring of New Delhi metallo-beta-lactamase activity in living bacterial cells by 1H NMR. Angew. Chemie Int. Ed. 53, 2130-2133