The development of new platform technologies for high-throughput screening of cellular targets will be a central goal of network. For example carbohydrate  arrays are being developed for studying carbohydrate-binding proteins and carbohydrate-processing enzymes (UK glycoarray network; ITN Marie Curie network EuroGlycoarrays; FP7 research programme GlycoBioM) (Flitsch). This platform provides a new approach to screen for modulators of cell adhesion, carbohydrate–antibody binding, and cell surface oligosaccharide biosynthesis which are implicated in many disease processes [1]. Semi-synthetic methods for the synthesis of defined glycoproteins to investigate the role that glycosylation plays within cells (Flitsch) [2] are also being applied to produce therapeutic proteins (antibodies, glycoprotein hormones etc. with Oxyrane).

Functional protein arrays are being developed to map protein phosphorylation, glycosyslation and ubiquitination and to screen for inhibitors of protein:protein and protein:nucleic acid interactions (Micklefield, Wong) [3]. This has involved the development of site-selective and covalent method to immobilise proteins for chip-based assays [3, 4].

Structural Chemical Biology. The network includes some of the UK’s leading structural chemical biologists. Notably, time resolved X-ray crystallography allows us to visualize in real time and with atomic detail the conformational changes of proteins on ligand or substrate binding [5]. We have developed novel neutron crystallography techniques to directly assign the positions of hydrogen isotopes in a protein and its bound solvent, providing more information than traditional X-ray diffraction (Helliwell) [6].

New NMR methodologies are essential in defining the structure and dynamics of cellular targets. We will use novel 19F NMR methods to probe the structure and mechanism of phosphoryl transfer enzymes, key to the development of new kinase inhibitors (Waltho) [7,8]. Isotopically discriminated NMR spectroscopy (IDIS-NMR) will be applied to study protein-protein interactions and to screen for ligands that modulate protein-protein interactions (Golovanov) [9]. In addition to traditional biophysical techniques such as ITC, SPR etc., we are exploring entirely novel methods to probe interactions between small-molecules and biomolecular targets. This includes Raman spectroscopy, which has been used to monitor protein folding (Goodacre, Blanch) [10] and along with Avacta, work is underway to design a new stopped-flow Raman spectrometer to probe kinetics and thermodynamics of drug/ligand binding with proteins and RNA targets.

Medicinal enzymology and biochemistry. Expertise in structural and mechanistic enzymology along with chemical biology, is allowing new enzyme targets to be validated and new inhibitors to be designed for vascular adhesion protein 1 (VAP-1) a target for autoimmune and chronic inflammatory conditions (Scrutton). Structure-based drug design is guiding the development of potent inhibitors of MptpB, a secreted phosphatase, essential for Mycobacterium tuberculosis virulence and survival in macrophages(Tabernero, 11, 12). Small molecule inhibitors are also used to probe the structure and mechanism of P450 targets in Mycobacterium tuberculosis, which will drive fragment based screening efforts to generate new leads for TB [13,14]. In addition to smaller enzymes, we are targeting large multi protein complexes such as the mammalian spliceosome (O’Keefe, Berrisford) as well as quorum sensing signaling pathways in bacteria (Freeman, Gardiner) [15].
 

Cell biology & Cancer Research. New chemical tools are being used to probe cell cycle regulation and in particular mitosis (Taylor). Indeed, we have a world-leading reputation in the area of mitotic inhibitors and were the first to develop inhibitors for Mps1 and aurora kinases [16, 17]. Novel small molecule modulators of the nuclear hormone receptors, glucocorticoid receptor and REV-ERBα are being applied to inflammation and to metabolic control (Ray & Loudon) [18], and collaboration with Pfizer will test the biological action of novel casein kinase inhibitors with a view to regulating the biological clock (Loudon)[19].

Selective inhibition of protein translocation at the endoplasmic reticulum, and the resulting accumulation of potentially cytotoxic precursors, is an area of major interest. Perturbation of ER translocation represents a novel mode of action for small molecules and is one we will pursue in combination with other small molecules that target the ubiquitin-proteasome system (High & Swanton) [20,21].

Cancer Research in Manchester is world renown, both within the University and the NHS. Through our links with the medical faculty (FHMS, Dive, Stratford, Ray) and the Paterson Institute (Oglivie, Jordan) the network has access to one of largest groupings of scientists and medics from the oncology field in the UK.

[1] Flitsch et al. Chem. Commun., 2008, 4400-4412 [2] Flitsch et al. FEBS J. 2010, 277, 2171; [3] Micklefield, Wong et al. Chem. Rev. 2009, 109, 4025; [4] Micklefield, Wong et al. J. Am. Chem. Soc. 2008, 130, 12456; [5] Leys et al. Science 2006, 312, 237; [6] Helliwell et al. Proc Nat. Acad. Sci. USA,  2004, 101, 16405; [7] Waltho et al. Proc Nat. Acad. Sci. USA 2010, 107, 4555;  [8] Waltho et al. J. Mol. Biol. 2010, 396, 345; [9] Golovanov et al. J. Amer. Chem. Soc. 2007, 129, 6528 [10] Blanch et al. Analytical Chemistry 2010, 82, 6347; [11] Munro et al. Arch Biochem Biophys. 2007, 464, 228; [12] Tabernero et al. Biochem. J. 2007,406, 13-18; [13] Tabernero et al. J Antimicrob Chemother. 2009, 63, 928-36.  [14] Munro et al. Biol. Chem. 2006, 281, 39437; [15] Freeman, Gardiner et al. Bioorg. Med. Chem. Lett. 2010, 20, 2625; [16] Taylor et al. J. Cell Biol. 2010, 190, 25;  Curr. Opin. Cell. Biol. 2008, 20, 77;  [17] Taylor et al. Nat Rev Cancer 2004, 4, 927-36; [18] Ray et al. Endocrinology 2009, 150, 75; [19] Loudon et al. Proc. Natl. Acad. Sci. USA 2010, 107, 15240; [20] High, Swanton et al. Proc. Natl. Acad. Sci. USA 2005, 102, 4342; [21] High, Swanton et al. J. Cell Sci. 2009, 122, 4393;