Publications
241 results found
Storck EM, Serwa RA, Tate EW, 2013, Chemical proteomics: a powerful tool for exploring protein lipidation, Biochemical Society Transactions, Vol: 41, Pages: 56-61, ISSN: 1470-8752
The study of post-translational modifications such as protein lipidation is a non-trivial challenge of the post-genomic era. In recent years the field of chemical proteomics has greatly advanced our ability to identify and quantify protein lipidation. In the present review, we give a brief overview of the tools available to study protein acylation, prenylation and cholesterylation, and their application in the identification and quantification of protein lipidation in health and disease.
Rackham MD, Brannigan JA, Moss DK, et al., 2013, Discovery of Novel and Ligand-Efficient Inhibitors of <i>Plasmodium falciparum</i> and <i>Plasmodium vivax N</i>-Myristoyltransferase, JOURNAL OF MEDICINAL CHEMISTRY, Vol: 56, Pages: 371-375, ISSN: 0022-2623
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- Citations: 51
Douse CH, Green JL, Salgado PS, et al., 2012, Regulation of the Plasmodium motor complex: phosphorylation of myosin A tail-interacting protein (MTIP) loosens its grip on MyoA., Journal of Biological Chemistry, Vol: 287, Pages: 36968-36977, ISSN: 1083-351X
Background: Recent phosphoproteome data reveal the extent of post-translational phosphorylation in selected apicomplexanparasites.Results: Binding site mutants that mimic the effect of MTIP phosphorylation in vivo severely decrease MyoA binding.Conclusion: Phosphorylation of selected binding site residues modulates the activity of the actomyosin motor.Significance: Study of Apicomplexa phosphosites can inform on the regulation of functions involved in pathogenesis.
Yu Z, Brannigan JA, Moss DK, et al., 2012, Design and Synthesis of Inhibitors of Plasmodium falciparum N-Myristoyltransferase, A Promising Target for Antimalarial Drug Discovery, Journal of Medicinal Chemistry, Vol: 55, Pages: 8879-8890, ISSN: 0022-2623
Design of inhibitors for N-myristoyltransferase (NMT), an enzyme responsible for protein trafficking in Plasmodium falciparum, the most lethal species of parasites that cause malaria, is described. Chemistry-driven optimization of compound 1 from a focused NMT inhibitor library led to the identification of two early lead compounds 4 and 25, which showed good enzyme and cellular potency and excellent selectivity over human NMT. These molecules provide a valuable starting point for further development.
Thinon E, Mann D, Tate EW, 2012, Targeting N-myristoyl transferase-1 in cancer using peptide microarrays, Journal of Peptide Science, Vol: 18, Pages: S75-S75, ISSN: 1099-1387
Price HP, Hodgkinson MR, Wright MH, et al., 2012, A role for the vesicle-associated tubulin binding protein ARL6 (BBS3) in flagellum extension in Trypanosoma brucei, Biochimica et Biophysica Acta-Molecular Cell Research, Vol: 1823, Pages: 1178-1191, ISSN: 0167-4889
The small GTPase Arl6 is implicated in the ciliopathic human genetic disorder Bardet–Biedl syndrome, actingat primary cilia in recruitment of the octomeric BBSome complex, which is required for specific traffickingevents to and from the cilium in eukaryotes. Here we describe functional characterisation of Arl6 in the flagellatedmodel eukaryote Trypanosoma brucei, which requires motility for viability. Unlike human Arl6 whichhas a ciliary localisation, TbARL6 is associated with electron-dense vesicles throughout the cell body followingco-translational modification by N-myristoylation. Similar to the related protein ARL-3A in T. brucei, modulationof expression of ARL6 by RNA interference does not prevent motility but causes a significant reductionin flagellum length. Tubulin is identified as an ARL6 interacting partner, suggesting that ARL6 may act as ananchor between vesicles and cytoplasmic microtubules. We provide evidence that the interaction betweenARL6 and the BBSome is conserved in unicellular eukaryotes. Overexpression of BBS1 leads to translocationof endogenous ARL6 to the site of exogenous BBS1 at the flagellar pocket. Furthermore, a combination ofBBS1 overexpression and ARL6 RNAi has a synergistic inhibitory effect on cell growth. Our findings indicatethat ARL6 in trypanosomes contributes to flagellum biogenesis, most likely through an interaction with theBBSome
Konitsiotis A, Chang S, Masumoto N, et al., 2012, Hhat a Potential New Target in Treatment of Pancreatic Cancer, 22nd Biennial Congress of the European-Association-for-Cancer-Research, Publisher: ELSEVIER SCI LTD, Pages: S149-S149, ISSN: 0959-8049
Bell AS, Mills JE, Williams GP, et al., 2012, Selective Inhibitors of Protozoan Protein N-myristoyltransferases as Starting Points for Tropical Disease Medicinal Chemistry Programs, PLOS Neglected Tropical Diseases, Vol: 6, ISSN: 1935-2735
Inhibition of N-myristoyltransferase has been validated pre-clinically as a target for the treatment of fungal andtrypanosome infections, using species-specific inhibitors. In order to identify inhibitors of protozoan NMTs, we chose toscreen a diverse subset of the Pfizer corporate collection against Plasmodium falciparum and Leishmania donovani NMTs.Primary screening hits against either enzyme were tested for selectivity over both human NMT isoforms (Hs1 and Hs2) andfor broad-spectrum anti-protozoan activity against the NMT from Trypanosoma brucei. Analysis of the screening results hasshown that structure-activity relationships (SAR) for Leishmania NMT are divergent from all other NMTs tested, a finding notpredicted by sequence similarity calculations, resulting in the identification of four novel series of Leishmania-selective NMTinhibitors. We found a strong overlap between the SARs for Plasmodium NMT and both human NMTs, suggesting thatachieving an appropriate selectivity profile will be more challenging. However, we did discover two novel series withselectivity for Plasmodium NMT over the other NMT orthologues in this study, and an additional two structurally distinctseries with selectivity over Leishmania NMT. We believe that release of results from this study into the public domain willaccelerate the discovery of NMT inhibitors to treat malaria and leishmaniasis. Our screening initiative is another example ofhow a tripartite partnership involving pharmaceutical industries, academic institutions and governmental/nongovernmentalorganisations such as Medical Research Council and Wellcome Trust can stimulate research for neglecteddiseases
Goncalves V, Brannigan JA, Whalley D, et al., 2012, Discovery of Plasmodium vivax N-Myristoyltransferase Inhibitors: Screening, Synthesis, and Structural Characterization of their Binding Mode, Journal of Medicinal Chemistry, Vol: 55, Pages: 3578-3582, ISSN: 0022-2623
N-Myristoyltransferase (NMT) is a prospective drug target against parasitic protozoa. Herein we report the successful discovery of a series of Plasmodium vivax NMT inhibitors by high-throughput screening. A high-resolution crystal structure of the hit compound in complex with NMT was obtained, allowing understanding of its novel binding mode. A set of analogues was designed and tested to define the chemical groups relevant for activity and selectivity.
Bradshaw RT, Aronica PGA, Tate EW, et al., 2012, Developing mutational locally enhanced sampling (MULES) for predicting relative binding free energies at protein-protein interfaces, 11th International Biorelated Polymer Symposium / 243rd National Spring Meeting of the American-Chemical-Society (ACS), Publisher: AMER CHEMICAL SOC, ISSN: 0065-7727
Goncalves V, Brannigan JA, Thinon E, et al., 2012, A fluorescence-based assay for N-myristoyltransferase activity, Analytical Biochemistry, Vol: 421, Pages: 342-344, ISSN: 1096-0309
N-myristoylation is the irreversible attachment of a C14 fatty acid, myristic acid, to the N-terminal glycine of a protein via formation of an amide bond. This modification is catalyzed by myristoyl–coenzyme A (CoA):protein N-myristoyltransferase (NMT), an enzyme ubiquitous in eukaryotes that is up-regulated in several cancers. Here we report a sensitive fluorescence-based assay to study the enzymatic activity of human NMT1 and NMT2 based on detection of CoA by 7-diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin. We also describe expression and characterization of NMT1 and NMT2 and assay validation with small molecule inhibitors. This assay should be broadly applicable to NMTs from a range of organisms.
Broncel M, Serwa RA, Tate EW, 2012, A New Chemical Handle for Protein AMPylation at the Host-Pathogen Interface, Chembiochem, Vol: 13, Pages: 183-185, ISSN: 1439-7633
agging protein AMPylation: A new chemical reporter for AMPylation, recently identified as a key post-translational modification during bacterial infection, is a robust tool for detecting and identifying AMPylated proteins in vitro.
Dang THT, Fagan RP, Fairweather NF, et al., 2012, Novel inhibitors of surface layer processing in Clostridium difficile, Bioorganic & Medicinal Chemistry, Vol: 20, Pages: 614-621, ISSN: 1464-3391
Clostridium difficile, a leading cause of hospital-acquired bacterial infection, is coated in a dense surface layer (S-layer) that is thought to provide both physicochemical protection and a scaffold for host-pathogen interactions. The key structural components of the S-layer are two proteins derived from a polypeptide precursor, SlpA, via proteolytic cleavage by the protease Cwp84. Here, we report the design, synthesis and in vivo characterization of a panel of protease inhibitors and activity-based probes (ABPs) designed to target S-layer processing in live C. difficile cells. Inhibitors based on substrate-mimetic peptides bearing a C-terminal Michael acceptor warhead were found to be promising candidates for further development.
Heal WP, Wright MH, Thinon E, et al., 2012, Multifunctional protein labeling via enzymatic N-terminal tagging and elaboration by click chemistry, NATURE PROTOCOLS, Vol: 7, Pages: 105-117, ISSN: 1754-2189
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- Citations: 82
Heal WP, Tate EW, 2012, Application of Activity-Based Protein Profiling to the Study of Microbial Pathogenesis, ACTIVITY-BASED PROTEIN PROFILING, Vol: 324, Pages: 115-135, ISSN: 0340-1022
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- Citations: 17
Furse S, Brooks NJ, Seddon AM, et al., 2012, Lipid membrane curvature induced by distearoyl phosphatidylinositol 4-phosphate, Soft Matter
Bradshaw RT, Aronica PGA, Tate EW, et al., 2012, Mutational Locally Enhanced Sampling (MULES) for quantitative prediction of the effects of mutations at protein-protein interfaces, CHEMICAL SCIENCE, Vol: 3, Pages: 1503-1511, ISSN: 2041-6520
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- Citations: 2
Tate EW, Goss RJM, 2011, Highlights from the 46th EUCHEM Conference on stereochemistry, Bürgenstock, Switzerland, May 2011., Chem Commun (Camb), Vol: 47, Pages: 10869-10873
Delmotte A, Tate EW, Yaliraki SN, et al., 2011, Protein multi-scale organization through graph partitioning and robustness analysis: application to the myosin-myosin light chain interaction, PHYSICAL BIOLOGY, Vol: 8, ISSN: 1478-3975
Despite the recognized importance of the multi-scale spatio-temporal organization of proteins, most computational tools can only access a limited spectrum of time and spatial scales, thereby ignoring the effects on protein behavior of the intricate coupling between the different scales. Starting from a physico-chemical atomistic network of interactions that encodes the structure of the protein, we introduce a methodology based on multi-scale graph partitioning that can uncover partitions and levels of organization of proteins that span the whole range of scales, revealing biological features occurring at different levels of organization and tracking their effect across scales. Additionally, we introduce a measure of robustness to quantify the relevance of the partitions through the generation of biochemically-motivated surrogate random graph models. We apply the method to four distinct conformations of myosin tail interacting protein, a protein from the molecular motor of the malaria parasite, and study properties that have been experimentally addressed such as the closing mechanism, the presence of conserved clusters, and the identification through computational mutational analysis of key residues for binding.
de la Riva L, Willing SE, Tate EW, et al., 2011, Roles of Cysteine Proteases Cwp84 and Cwp13 in Biogenesis of the Cell Wall of <i>Clostridium difficile</i>, JOURNAL OF BACTERIOLOGY, Vol: 193, Pages: 3276-3285, ISSN: 0021-9193
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- Citations: 42
Serwa R, Tate EW, 2011, Activity-based profiling for drug discovery, Chemistry and Biology, Vol: 18, Pages: 407-409, ISSN: 1879-1301
Activity-based protein profiling (ABPP) is emerging as a game-changing tool for drug discovery, target validation, and basic biology. In this issue, Chang et al. (2011) report the ABPP-facilitated discovery of JW480, a highly selective potent and orally bioavailable inhibitor of monoalkylglycerol ether hydrolase KIAA1363 that dramatically impairs in vivo growth of human prostate cancer cell lines.
Heal WP, Dang TH, Tate EW, 2011, Activity-based probes: discovering new biology and new drug targets., Chem Soc Rev, Vol: 40, Pages: 246-257, ISSN: 1460-4744
The development and application of chemical technologies enabling direct analysis of enzyme activity in living systems has undergone explosive growth in recent years. Activity-based protein profiling (ABPP) is a key constituent of this broad field, and is among the most powerful and mature chemical proteomic technologies. This tutorial review introduces the essential features of ABPP and the design and application of activity-based probes (ABPs) from drug target elucidation and in vivo visualisation of enzyme activity to comprehensive profiling of the catalytic content of living systems, and the discovery of new biological pathways.
Heal WP, Jovanovic B, Bessin S, et al., 2011, Bioorthogonal chemical tagging of protein cholesterylation in living cells, CHEMICAL COMMUNICATIONS, Vol: 47, Pages: 4081-4083, ISSN: 1359-7345
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- Citations: 69
Bradshaw RT, Patel BH, Tate EW, et al., 2010, Comparing experimental and computational alanine scanning techniques for probing a prototypical protein-protein interaction, PROTEIN ENGINEERING DESIGN & SELECTION, Vol: 24, Pages: 197-207, ISSN: 1741-0126
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- Citations: 62
Berry AFH, Heal WP, Tarafder AK, et al., 2010, Rapid Multilabel Detection of Geranylgeranylated Proteins by Using Bioorthogonal Ligation Chemistry, CHEMBIOCHEM, Vol: 11, Pages: 771-773, ISSN: 1439-4227
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- Citations: 39
Brannigan JA, Smith BA, Yu Z, et al., 2010, N-Myristoyltransferase from Leishmania donovani: Structural and Functional Characterisation of a Potential Drug Target for Visceral Leishmaniasis, Journal of Molecular Biology, Vol: 396, Pages: 985-999, ISSN: 1089-8638
N-Myristoyltransferase (NMT) catalyses the attachment of the 14-carbon saturated fatty acid, myristate, to the amino-terminal glycine residue of a subset of eukaryotic proteins that function in multiple cellular processes, including vesicular protein trafficking and signal transduction. In these pathways, N-myristoylation facilitates association of substrate proteins with membranes or the hydrophobic domains of other partner peptides. NMT function is essential for viability in all cell types tested to date, demonstrating that this enzyme has potential as a target for drug development. Here, we provide genetic evidence that NMT is likely to be essential for viability in insect stages of the pathogenic protozoan parasite, Leishmania donovani, causative agent of the tropical infectious disease, visceral leishmaniasis. The open reading frame of L. donovaniNMT has been amplified and used to overproduce active recombinant enzyme in Escherichia coli, as demonstrated by gel mobility shift assays of ligand binding and peptide-myristoylation activity in scintillation proximity assays. The purified protein has been crystallized in complex with the non-hydrolysable substrate analogue S-(2-oxo)pentadecyl-CoA, and its structure was solved by molecular replacement at 1.4 Å resolution. The structure has as its defining feature a 14-stranded twisted β-sheet on which helices are packed so as to form an extended and curved substrate-binding groove running across two protein lobes. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the carboxy-terminal lobe. These studies validate L. donovani NMT as a potential target for development of new therapeutic agents against visceral leishmaniasis.
Dang THT, de la Riva L, Fagan RP, et al., 2010, Chemical Probes of Surface Layer Biogenesis in <i>Clostridium difficile</i>, ACS CHEMICAL BIOLOGY, Vol: 5, Pages: 279-285, ISSN: 1554-8929
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- Citations: 48
Wright MH, Heal WP, Mann DJ, et al., 2010, Protein myristoylation in health and disease., J Chem Biol, Vol: 3, Pages: 19-35, ISSN: 1864-6166
N-myristoylation is the attachment of a 14-carbon fatty acid, myristate, onto the N-terminal glycine residue of target proteins, catalysed by N-myristoyltransferase (NMT), a ubiquitous and essential enzyme in eukaryotes. Many of the target proteins of NMT are crucial components of signalling pathways, and myristoylation typically promotes membrane binding that is essential for proper protein localisation or biological function. NMT is a validated therapeutic target in opportunistic infections of humans by fungi or parasitic protozoa. Additionally, NMT is implicated in carcinogenesis, particularly colon cancer, where there is evidence for its upregulation in the early stages of tumour formation. However, the study of myristoylation in all organisms has until recently been hindered by a lack of techniques for detection and identification of myristoylated proteins. Here we introduce the chemistry and biology of N-myristoylation and NMT, and discuss new developments in chemical proteomic technologies that are meeting the challenge of studying this important co-translational modification in living systems.
So S, Peeva LG, Tate EW, et al., 2010, Organic Solvent Nanofiltration: A New Paradigm in Peptide Synthesis, Org Process Res Dev, Vol: 14, Pages: 1313-1325
Thomas JC, Green JL, Howson RI, et al., 2010, Interaction and dynamics of the <i>Plasmodium</i> <i>falciparum</i> MTIP-MyoA complex, a key component of the invasion motor in the malaria parasite, MOLECULAR BIOSYSTEMS, Vol: 6, Pages: 494-498, ISSN: 1742-206X
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- Citations: 25
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