A5014189721- Enzyme Structure and Function
- Cellular transport and secretion
- Glycosylation and Glycoproteins Research
- RNA and protein synthesis mechanisms
- SARS-CoV-2 and COVID-19 Research
- Microbial Metabolites in Food Biotechnology
- Microtubule and mitosis dynamics
- Porphyrin Metabolism and Disorders
- Lysosomal Storage Disorders Research
- Retinal Development and Disorders
- Protein Kinase Regulation and GTPase Signaling
- Plant-Microbe Interactions and Immunity
- Neurological diseases and metabolism
- SARS-CoV-2 detection and testing
- COVID-19 Clinical Research Studies
- CRISPR and Genetic Engineering
- Biochemical and Molecular Research
- Parkinson's Disease Mechanisms and Treatments
- Carbohydrate Chemistry and Synthesis
- Fluorine in Organic Chemistry
- Microbial Natural Products and Biosynthesis
- Advanced Proteomics Techniques and Applications
- Sphingolipid Metabolism and Signaling
- Dendrimers and Hyperbranched Polymers
- Genetics and Neurodevelopmental Disorders
University of Dundee
2010-2024
MRC Protein Phosphorylation and Ubiquitylation Unit
2019-2021
Medical Research Council
2020-2021
University of St Andrews
2003-2010
The recent emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the underlying cause Disease 2019 (COVID-19), has led to a worldwide pandemic causing substantial morbidity, mortality, and economic devastation. In response, many laboratories have redirected attention SARS-CoV-2, meaning there is an urgent need for tools that can be used in unaccustomed working with coronaviruses. Here we report range SARS-CoV-2 research. First, describe facile single plasmid reverse...
Mutations that activate LRRK2 protein kinase cause Parkinson's disease. phosphorylates a subset of Rab GTPases within their Switch-II motif controlling interaction with effectors. An siRNA screen all human phosphatases revealed poorly studied phosphatase, PPM1H, counteracts signaling by specifically dephosphorylating proteins. PPM1H knockout increased endogenous phosphorylation and inhibited dephosphorylation in A549 cells. Overexpression suppressed LRRK2-mediated phosphorylation. also...
The Scottish Structural Proteomics Facility was funded to develop a laboratory scale approach high throughput structure determination. effort successful in that over 40 structures were determined. These and the methods harnessed obtain them are reported here. This report reflects on value of automation but also continued requirement for degree scientific technical expertise. efficiency process poses challenges current paradigm structural analysis publication. In 5 year period we published...
Autosomal dominant mutations in LRRK2 that enhance kinase activity cause Parkinson's disease. phosphorylates a subset of Rab GTPases including Rab8A and Rab10 within its effector binding motif. Here, we explore whether LRRK1, less studied homolog regulates growth factor receptor trafficking osteoclast biology might also phosphorylate proteins. Using mass spectrometry, found LRRK1 knock-out cells, phosphorylation Rab7A at Ser72 was most impacted. This residue lies the equivalent site targeted...
Abstract WNK family kinases are regulated by osmotic stress and control ion homeostasis activating SPAK OXSR1 kinases. Using a proximity ligation approach, we found that promotes the association of WNK1 with NRBP1 pseudokinase TSC22D2/4 adaptor proteins. is closely related to isoforms also contains RΦ-motif binding conserved C-terminal (CCT) domain, similar CCT domains in WNKs, OXSR1. Knockdown markedly inhibited sorbitol-induced activation downstream components. AlphaFold-3 modelling...
Chlorination of natural products is often required for their biological activity; notable examples include vancomycin, the last-ditch antibiotic. It now known that many chlorinated are made not by haloperoxidases, but FADH2-dependent halogenases. The mechanism flavin-containing enzymes obscure and there no structural data. Here, crystals PrnA (tryptophan 7-halogenase), an enzyme regioselectively chlorinates tryptophan, cocrystallized with tryptophan FAD reported. belong to tetragonal space...
Ranasmurfin, a previously uncharacterized ∼13 kDa blue protein found in the nests of frog Polypedates leucomystax, has been purified and crystallized. The crystals are an intense colour diffract to 1.51 Å with P21 symmetry unit-cell parameters = 40.9, b 59.9, c 45.0 Å, β 93.3°. Self-rotation function analysis indicates presence dimer asymmetric unit. Biochemical data suggest that is related formation. Sequence for incomplete, but thus far have identified no model molecular replacement. A...
Organofluorine compounds are widely prepared throughout the chemicals industry, but their prepararion generally requires harsh fluorinating reagents and non-aqueous solvents. On other hand, biology has hardly exploited organofluorine compounds. A very few organisms synthesize metabolites, suggesting they have evolved a mechanism to overcome kinetic desolvation barrier utilizing F−(aq). Here, purification crystallization of an enzyme from Streptomyces cattleya which is responsible for...
Sar2676, a pantothenate synthetase with molecular weight of 31 419 Da from methicillin-resistant Staphylococcus aureus, has been expressed, purified and crystallized at 293 K. The protein crystallizes in primitive triclinic lattice, unit-cell parameters = 45.3, b 60.5, c 117.6 Å, α 87.2, β 81.2, γ 68.4°. A complete data set collected to 2.3 Å resolution the ESRF. Consideration likely solvent content suggested asymmetric unit contain four molecules. This confirmed by molecular-replacement...
Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase with a molecular weight of 48,168 Da, was overexpressed in methicillin-resistant Staphylococcus aureus compared methicillin-sensitive strain. The protein expressed Escherichia coli, purified and crystallized. crystallized primitive orthorhombic Laue group unit-cell parameters = 83.6, b 91.3, c 106.0 A, alpha beta gamma 90 degrees. Analysis the systematic absences along three principal axes...
Abstract Mutations that activate LRRK2 protein kinase cause Parkinson’s disease. phosphorylates a subset of Rab GTPases within their Switch-II motif controlling interaction with effectors. An siRNA screen all phosphatases revealed poorly studied phosphatase, PPM1H, counteracts signaling by specifically dephosphorylating proteins. PPM1H knock out increased endogenous phosphorylation and inhibited dephosphorylation. Overexpression suppressed LRRK2-mediated phosphorylation. also efficiently...
Abstract Much attention has focused on LRRK2, as autosomal dominant missense mutations that enhance its kinase activity cause inherited Parkinson’s disease. LRRK2 regulates biology by phosphorylating a subset of Rab GTPases including Rab8A and Rab10 within effector binding motif. In this study we explore whether LRRK1, less studied homologue growth factor receptor trafficking osteoclast might also phosphorylate proteins. Using mass spectrometry, found the endogenous Rab7A protein,...
Sso6206, a 10.5 kDa protein from Sulfolobus solfataricus, has been overexpressed, purified and crystallized. The crystallizes in space group P61/522, with unit-cell parameters = b 157.8, c 307.3 Å. crystals are hexagonal bipyramids data set collected to 2.4 Å resolution. Molecular replacement cannot be attempted as no convincing model can identified. Crystals of selenomethionine-variant have not yet obtained. Interestingly, crystal packing, gel filtration mass spectrometry all suggest the...