A5005716688- Pharmacogenetics and Drug Metabolism
- Tuberculosis Research and Epidemiology
- Metal-Catalyzed Oxygenation Mechanisms
- Biochemical and Molecular Research
- Computational Drug Discovery Methods
- Microbial Natural Products and Biosynthesis
- Chemical Reactions and Isotopes
- Cancer therapeutics and mechanisms
- Steroid Chemistry and Biochemistry
- Porphyrin Metabolism and Disorders
- Mycobacterium research and diagnosis
- Cancer Treatment and Pharmacology
- Microbial Metabolic Engineering and Bioproduction
- Heme Oxygenase-1 and Carbon Monoxide
- Folate and B Vitamins Research
- Hemoglobin structure and function
- Enzyme Structure and Function
- Biosimilars and Bioanalytical Methods
- Analytical Chemistry and Chromatography
- Drug Transport and Resistance Mechanisms
- Microbial bioremediation and biosurfactants
- Synthesis and Reactions of Organic Compounds
- Antibiotic Resistance in Bacteria
- Innovative Microfluidic and Catalytic Techniques Innovation
- Hormonal Regulation and Hypertension
University of Manchester
2012-2024
University of Huddersfield
2021-2023
Salford Royal NHS Foundation Trust
2011
Manchester Academic Health Science Centre
2011
Baoshan College
2010
Piaggio (Italy)
2009
University of Leicester
2002-2007
Weatherford College
2007
La Trobe University
2007
University of East Anglia
2006
Cytochrome P450 monooxygenases play a crucial role in the biosynthesis of many natural products and human metabolism numerous pharmaceuticals. This has inspired synthetic organic medicinal chemists to exploit them as catalysts regio- stereoselective CH-activating oxidation structurally simple complex compounds such steroids. However, levels stereoselectivity well activity are not routinely high enough for real applications. Protein engineering using rational design or directed evolution...
The cholesterol-lowering blockbuster drug pravastatin can be produced by stereoselective hydroxylation of the natural product compactin. We report here metabolic reprogramming antibiotics producer Penicillium chrysogenum toward an industrial production process. Following successful introduction compactin pathway into β-lactam-negative P. DS50662, a new cytochrome P450 (P450 or CYP) from Amycolatopsis orientalis (CYP105AS1) was isolated to catalyze final step. Structural and biochemical...
The genome sequence of Mycobacterium tuberculosis has revealed the presence 20 different cytochrome P450 mono-oxygenases (P450s) within this organism, and subsequent sequences other mycobacteria Streptomyces coelicolor have indicated that these actinomycetes also large complements P450s, pointing to important physiological roles for enzymes. actinomycete P450s include homologues 14α-sterol demethylases, targets azole class drugs in yeast fungi. Previously, type was considered be absent from...
The first structure of a P450 to an atomic resolution 1.06 Å has been solved for CYP121 from<i>Mycobacterium tuberculosis</i>. A comparison with EryF (CYP107A1) reveals remarkable overall similarity in fold major differences residing active site structural elements. high obtained allows visualization several unusual aspects. heme cofactor is bound two distinct conformations while being notably kinked one pyrrole group due close interaction the proline residue (Pro<sup>346</sup>) immediately...
Mycobacterium tuberculosis (Mtb) cytochrome P450 gene CYP121 is shown to be essential for viability of the bacterium in vitro by knock-out with complementation. Production protein Mtb cells demonstrated. Minimum inhibitory concentration values azole drugs against H37Rv were determined, rank order which correlated well Kd their binding CYP121. Solution-state spectroscopic, kinetic, and thermodynamic studies crystal structure determination a series active site mutants provide further insights...
Azole and triazole drugs are cytochrome P450 inhibitors widely used as fungal antibiotics possessing potent antimycobacterial activity. We present here the crystal structure of Mycobacterium tuberculosis CYP121 in complex with drug fluconazole, revealing a new azole heme ligation mode. In contrast to other structurally characterized complexes, where nitrogen directly coordinates iron, fluconazole does not displace aqua sixth ligand but occupies position that allows formation direct hydrogen...
We report characterization and the crystal structure of Mycobacterium tuberculosis cytochrome P450 CYP125, a implicated in metabolism host cholesterol essential for establishing infection mice. CYP125 is purified high spin form undergoes both type I II spectral shifts with various azole drugs. The 1.4-Å ligand-free reveals "letterbox" active site cavity dimensions appropriate entry polycyclic sterol. A mixture hexa-coordinate penta-coordinate states could be discerned, water binding as 6th...
The <i>Mycobacterium tuberculosis</i> cytochrome P450 enzyme CYP142 is encoded in a large gene cluster involved metabolism of host cholesterol. was expressed and purified as soluble, low spin hemoprotein. binds tightly to cholesterol its oxidized derivative cholest-4-en-3-one, with extensive shift the heme iron high state. High affinity for azole antibiotics demonstrated, highlighting their therapeutic potential. catalyzes either 27-hydroxylation cholesterol/cholest-4-en-3-one or generates...
To examine how azole inhibitors interact with the heme active site of cytochrome P450 enzymes, we have performed a series density functional theory studies on binding. These are first interactions center and give fundamental insight into azoles inhibit catalytic function enzymes. Since come in many varieties, tested three typical motifs representing broad range azole-type inhibitors: methylimidazolate, methyltriazolate, pyridine. structural represent azoles, such as econazole, fluconazole,...
Pieces of the puzzle: The first fragment-based approach was used to target cytochrome P450 enzymes (CYPs) for drug development (see scheme). experiments provide new insights into binding site essential Mycobacterium tuberculosis CYP121 enzyme, and resulted in a promising novel lead compound based on fragment merging. Detailed facts importance specialist readers are published as "Supporting Information". Such documents peer-reviewed, but not copy-edited or typeset. They made available...
The essential enzyme CYP121 is a target for drug development against antibiotic resistant strains of Mycobacterium tuberculosis. A triazol-1-yl phenol fragment 1 was identified to bind using cascade biophysical assays. Synthetic merging and optimization produced 100-fold improvement in binding affinity, yielding lead compound 2 (KD = 15 μM). Deconstruction into its component retrofragments allowed the group efficiency structural motifs be assessed, identification more LE scaffolds...
Given the frequent use of DMSO in biochemical and biophysical assays, it is desirable to understand influence concentration on dissociation or unfolding behavior proteins. In this study, effects structure interactions avidin Mycobacterium tuberculosis (Mtb) CYP142A1 were assessed through collision-induced (CID) (CIU) as monitored by nanoelectrospray ionization–ion mobility–mass spectrometry (nESI-IM-MS). concentrations higher than 4% (v/v) destabilize tetramer toward unfolding, via both its...