A5015450695- TGF-β signaling in diseases
- Pulmonary Hypertension Research and Treatments
- Porphyrin Metabolism and Disorders
- Biochemical and Structural Characterization
- Biochemical and Molecular Research
- Enzyme Structure and Function
- Heterotopic Ossification and Related Conditions
- Viral Infectious Diseases and Gene Expression in Insects
- Microbial bioremediation and biosurfactants
- Neurobiology and Insect Physiology Research
- Silk-based biomaterials and applications
- Virus-based gene therapy research
- Connective Tissue Growth Factor Research
- Microbial metabolism and enzyme function
- Click Chemistry and Applications
- Peptidase Inhibition and Analysis
- Ubiquitin and proteasome pathways
- Folate and B Vitamins Research
- Viral gastroenteritis research and epidemiology
- Caveolin-1 and cellular processes
- Bioactive natural compounds
- Cerebrovascular and genetic disorders
- Monoclonal and Polyclonal Antibodies Research
- Biotin and Related Studies
- Polysaccharides Composition and Applications
University of Cambridge
2018-2023
Papworth Hospital
2023
Lung Institute
2022
University College London
2013-2021
Cambridge School
2018-2021
Aberystwyth University
2016
University of the Punjab
2016
The Royal Free Hospital
2013-2015
Covalent probes can display unmatched potency, selectivity, and duration of action; however, their discovery is challenging. In principle, fragments that irreversibly bind target overcome the low affinity limits reversible fragment screening, but such electrophilic were considered nonselective rarely screened. We hypothesized mild electrophiles might selectivity challenge constructed a library 993 mildly fragments. characterized this by new high-throughput thiol-reactivity assay screened...
Rationale: Recently, rare heterozygous mutations in GDF2 were identified patients with pulmonary arterial hypertension (PAH). encodes the circulating BMP (bone morphogenetic protein) type 9, which is a ligand for BMP2 receptor.Objectives: Here we determined functional impact of and characterized plasma BMP9 BMP10 levels idiopathic PAH.Methods: Missense mutant proteins expressed vitro on protein processing secretion, endothelial signaling, activity was assessed. Plasma assayed PAH variants...
Abstract Activin receptor-like kinase 1 (ALK1)-mediated endothelial cell signalling in response to bone morphogenetic protein 9 (BMP9) and BMP10 is of significant importance cardiovascular disease cancer. However, detailed molecular mechanisms ALK1-mediated remain unclear. Here, we report crystal structures the BMP10:ALK1 complex at 2.3 Å prodomain-bound BMP9:ALK1 3.3 Å. Structural analyses reveal a tripartite recognition mechanism that defines BMP9 specificity for ALK1, predict...
L-Asparaginases catalyse the hydrolysis of asparagine to aspartic acid and ammonia. In addition, L-asparaginase is involved in biosynthesis amino acids such as lysine, methionine threonine. These enzymes have been used chemotherapeutic agents for treatment acute lymphoblastic leukaemia other haematopoietic malignancies since tumour cells cannot synthesize sufficient L-asparagine are thus killed by deprivation this acid. also food industry potential development biosensors, example levels...
Rationale: Autoimmunity is believed to play a role in idiopathic pulmonary arterial hypertension (IPAH). It not clear whether this causative or bystander of disease and if it carries any prognostic treatment significance. Objectives: To study autoimmunity IPAH using large cross-sectional cohort. Methods: Assessment the circulating immune cell phenotype was undertaken flow cytometry, profile serum immunoglobulins generated standardized multiplex array 19 clinically validated autoantibodies...
A number of X-ray analyses an enzyme involved in a key early stage tetrapyrrole biosynthesis are reported. Two structures human 5-aminolaevulinate dehydratase (ALAD), native and recombinant, have been determined at 2.8 Å resolution, showing that the adopts octameric quaternary structure accord with previously published from range other species. However, this is contrast to finding disease-related F12L mutant uniquely forms hexamers [Breinig et al. (2003), Nature Struct. Biol. 10 , 757–763]....
Abstract Bone morphogenetic proteins (BMPs) are multifunctional cytokines of the transforming growth factor β (TGFβ) superfamily with potential therapeutic applications due to their broad biological functionality. Designing BMP mimetics specific activity will contribute translational BMP-based therapies. Here, we report a BMP9 peptide mimetic, P3, designed from type I receptor binding site, which showed millimolar affinities for activin like kinase 1 (ALK1), ALK2 and ALK3. Although showing...
Abstract Heterozygous mutations in BMPR2 (bone morphogenetic protein (BMP) receptor type II) cause pulmonary arterial hypertension. BMPRII is a for over 15 BMP ligands, but why lung-specific pathology unknown. To elucidate the molecular basis of BMP:BMPRII interactions, we report crystal structures binary and ternary complexes with BMP10, which contain an ensemble seven different BMP10:BMPRII 1:1 complexes. binds BMP10 at knuckle epitope, A-loop β4 strand making BMPRII-specific interactions....
Abstract Covalent probes can display unmatched potency, selectivity and duration of action, however, their discovery is challenging. In principle, fragments that irreversibly bind target overcome the low affinity limits reversible fragment screening. Such electrophilic were considered non-selective rarely screened. We hypothesized mild electrophiles might challenge, constructed a library 993 mildly fragments. characterized this by new high-throughput thiol-reactivity assay screened them...
During efforts to crystallize the enzyme 2,4-dihydroxyacetophenone dioxygenase (DAD) from Alcaligenes sp. 4HAP, a small number of strongly diffracting protein crystals were obtained after two years crystal growth in one condition. The diffracted synchrotron radiation almost 1.0 Å resolution and were, until recently, assumed be formed by DAD protein. However, when another form this was eventually solved at lower resolution, molecular replacement using new structure as search model did not...
Pullulan-hydrolysing enzymes, more commonly known as debranching enzymes for starch and other polysaccharides, are of great interest have been widely used in the starch-saccharification industry. Type III pullulan hydrolase from Thermococcus kodakarensis (TK-PUL) possesses both pullulanase α-amylase activities. Until now, only two this class, which capable hydrolysing α-1,4- α-1,6-glycosidic bonds to produce a mixture maltose, panose maltotriose, described. TK-PUL shows highest activity...
The enzyme 2,4′-dihydroxyacetophenone dioxygenase (DAD) catalyses the conversion of to 4-hydroxybenzoic acid and formic with incorporation molecular oxygen. Whilst vast majority dioxygenases cleave within aromatic ring substrate, DAD is very unusual in that it involved C—C bond cleavage a substituent ring. There evidence homotetramer 20.3 kDa subunits, each containing nonhaem iron, its sequence suggests belongs cupin family dioxygenases. In this paper, first X-ray structure from...
The enzyme porphobilinogen deaminase (PBGD) is one of the key enzymes in tetrapyrrole biosynthesis. It catalyses formation a linear from four molecules substrate (PBG). has dipyrromethane cofactor (DPM) active site which covalently linked to conserved cysteine residue through thioether bridge. are stepwise head-to-tail manner during reaction, catalysed by aspartate residue: Asp82 B. megaterium enzyme. Three mutations have been made affecting (D82A, D82E and D82N) their crystal structures...
Artocarpus incisa (breadfruit) seeds contain three different lectins (Frutalin, Frutapin (FTP) and Frutackin) with distinct carbohydrate specificities. The most abundant lectin is Frutalin, an α-D-galactose-specific carbohydrate-binding glycoprotein antitumour properties potential for tumour biomarker discovery as already reported. FTP the second abundant, but proved difficult to purify very low yields contamination Frutalin frustrating its characterization. Here, we report first time...
The enzyme 2,4′-dihydroxyacetophenone dioxygenase (DAD) catalyses the conversion of to 4-hydroxybenzoic acid and formic acid. This is a very unusual in that it cleaves C—C bond substituent aromatic ring rather than within itself. Whilst has been shown DAD tetramer solution, recently solved crystal structure Alcaligenes sp. 4HAP was fact dimeric tetrameric. Since use limited chymotrypsinolysis, which apparently results removal first 20 or so N-terminal residues DAD, necessary for...
The family B DNA polymerase from Pyrobaculum calidifontis ( Pc -polymerase) consists of 783 amino acids and is magnesium-ion dependent. It has an optimal pH 8.5, temperature 75°C a half-life 4.5 h at 95°C, giving it greater thermostability than the widely used Taq polymerase. enzyme also capable PCR-amplifying larger fragments up to 7.5 kb in length. was shown have functional, error-correcting 3′–5′ exonuclease activity, as do related high-fidelity polymerases Pyrococcus furiosus ,...
Outbreaks of human epidemic nonbacterial gastroenteritis are mainly caused by noroviruses. Viral replication requires a 3C-like cysteine protease (3CLpro) which processes the 200 kDa viral polyprotein into six functional proteins. The 3CLpro has attracted much interest due to its potential as target for antiviral drugs. A system growing high-quality crystals native Southampton norovirus (SV3CP) been established, allowing ligand-free crystal structure be determined 1.3 Å in tetrameric state....
Calexcitin was first identified in the marine snail Hermissenda crassicornis as a neuronal-specific protein that becomes upregulated and phosphorylated associative learning. possesses four EF-hand motifs, but only three (EF-1 to EF-3) are involved binding metal ions. Past work has indicated under physiological conditions EF-1 EF-2 bind Mg 2+ Ca , while EF-3 is likely . The fourth nonfunctional owing lack of key metal-binding residues. aim this study use crystallographic approach determine...
In many prokaryotes, the first step of threonine metabolism is catalysed by enzyme dehydrogenase (TDH), which uses NAD + to oxidize its substrate 2-amino-3-ketobutyrate. The absence a functional TDH gene in humans suggests that inhibitors this may have therapeutic potential against pathogens are reliant on enzyme. Here, from Clostridium difficile has been cloned and overexpressed, X-ray structure apoenzyme form determined at 2.6 Å resolution.