8 results on '"Owens, Ray"'
Search Results
2. In vitro and in vivo characterization of the multiple isoforms of Schistosoma mansoni hypoxanthine-guanine phosphoribosyltransferases.
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Romanello, Larissa, Zeraik, Ana Eliza, de Freitas Fernandes, Adriano, Torini, Juliana Roberta, Bird, Louise E., Nettleship, Joanne E., Rada, Heather, Reddivari, Yamini, Owens, Ray J., Serrão, Vitor Hugo Balasco, DeMarco, Ricardo, Brandão-Neto, José, and Pereira, Humberto D'Muniz
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SCHISTOSOMA mansoni , *CRYSTAL structure , *WASTE salvage - Abstract
Highlights • A crystal structure of Schistosoma mansoni HGPRT-1 (Sm HGPRT), complexed with IMP at a resolution of 2.8 Ǻ is described. • RNA-Seq and WISH data, suggests that some HGPRT isoforms might be involved in sexual maturation and reproduction in worms. • S. mansoni has four HGPRT isoforms, here we described the kinetics assays for three of them. Abstract Schistosoma mansoni , the parasite responsible for schistosomiasis, lacks the " de novo " purine biosynthetic pathway and depends entirely on the purine salvage pathway for the supply of purines. Numerous reports of praziquantel resistance have been described, as well as stimulated efforts to develop new drugs against schistosomiasis. Hypoxanthine-guanine phosphoribosyltransferase (HGPRT) is a key enzyme of the purine salvage pathway. Here, we describe a crystallographic structure of the S. mansoni HPGRT-1 (Sm HGPRT), complexed with IMP at a resolution of 2.8 Ǻ. Four substitutions were identified in the region of the active site between Sm HGPRT-1 and human HGPRT. We also present data from RNA-Seq and WISH, suggesting that some isoforms of HGPRT might be involved in the process related to sexual maturation and reproduction in worms; furthermore, its enzymatic assays show that the isoform Sm HGPRT-3 does not present the same catalytic efficiency as other isoforms. Finally, although other studies have previously suggested this enzyme as a potential antischistosomal chemotherapy target, the kinetics parameters reveal the impossibility to use Sm HGPRT as an efficient chemotherapeutic target. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
3. Structural and kinetic analysis of Schistosoma mansoni Adenylosuccinate Lyase (SmADSL).
- Author
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Romanello, Larissa, Serrão, Vitor Hugo Balasco, Torini, Juliana Roberta, Bird, Louise E., Nettleship, Joanne E., Rada, Heather, Reddivari, Yamini, Owens, Ray J., DeMarco, Ricardo, Brandão-Neto, José, and Pereira, Humberto D’Muniz
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SCHISTOSOMA mansoni , *LYASE genetics , *DRUG resistance , *TARGETED drug delivery , *PHARMACOKINETICS , *THERAPEUTICS - Abstract
Schistosoma mansoni is the parasite responsible for schistosomiasis, a disease that affects about 218 million people worldwide. Currently, both direct treatment and disease control initiatives rely on chemotherapy using a single drug, praziquantel. Concerns over the possibility of resistance developing to praziquantel, have stimulated efforts to develop new drugs for the treatment of schistosomiasis. Schistosomes do not have the de novo purine biosynthetic pathway, and instead depend entirely on the purine salvage pathway to supply its need for purines. The purine salvage pathway has been reported as a potential target for developing new drugs against schistosomiasis. Adenylosuccinate lyase ( Sm ADSL) is an enzyme in this pathway, which cleaves adenylosuccinate (ADS) into adenosine 5′-monophosphate (AMP) and fumarate. SmADSL kinetic characterization was performed by isothermal titration calorimetry (ITC) using both ADS and SAICAR as substrates. Structures of Sm ADSL in Apo form and in complex with AMP were elucidated by x-ray crystallography revealing a highly conserved tetrameric structure required for their function since the active sites are formed from residues of three different subunits. The active sites are also highly conserved between species and it is difficult to identify a potent species-specific inhibitor for the development of new therapeutic agents. In contrast, several mutagenesis studies have demonstrated the importance of dimeric interface residues in the stability of the quaternary structure of the enzyme. The lower conservation of these residues between Sm ADSL and human ADSL could be used to lead the development of anti-schistosomiasis drugs based on disruption of subunit interfaces. These structures and kinetics data add another layer of information to Schistosoma mansoni purine salvage pathway. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
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4. Some lessons from the systematic production and structural analysis of soluble αβ T-cell receptors
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van Boxel, Gijs I., Stewart-Jones, Guillaume, Holmes, Samantha, Sainsbury, Sarah, Shepherd, Dawn, Gillespie, G.M.A., Harlos, Karl, Stuart, David I., Owens, Ray, and Jones, E. Yvonne
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T cells , *CELL receptors , *MEMBRANE proteins , *CELLULAR immunity , *STRUCTURAL analysis (Science) , *MAJOR histocompatibility complex , *CRYSTALLIZATION , *ANTIGENS , *X-ray crystallography - Abstract
Abstract: T-cell receptors (TCRs) are membrane proteins which recognize antigens with high specificity forming the basis of the cellular immune response. The study of these receptors has been limited by the challenges in expressing sufficient quantities of stable soluble protein. Here we report our systematic approach for generating soluble, αβ-TCRs, for X-ray crystallographic studies. By using small-scale expression screens, novel standardized quality control mechanisms and crystallization and imaging robots we were able to add significantly to the current TCR structural database. Our success in crystallizing both isolated TCRs and Major histocompatibility complex (MHC):TCR complexes has provided us with sufficient data to develop focused crystallization screens, which have proved generically useful for the crystallization of this family of proteins and complexes. [Copyright &y& Elsevier]
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- 2009
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5. Novel nucleotide triphosphates as potent P2Y2 agonists with enhanced stability over UTP
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Davenport, Richard J., Diaz, Paloma, Galvin, Frances C.A., Lloyd, Steve, Mack, Stephen R., Owens, Ray, Sabin, Verity, and Wynn, Joanne
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NUCLEOTIDES , *PHOSPHATES , *LIGANDS (Biochemistry) , *ORGANIC compounds , *METABOLIC regulation - Abstract
Abstract: The synthesis of a series of novel C-linked nucleotide triphosphates is reported. These exhibit excellent agonist potency and selectivity for the P2Y2 receptor with a number of examples having EC50 values below 10nM. Representative compounds from the N-linked and C-linked series showed enhanced metabolic stability compared with that of the natural ligand UTP. [Copyright &y& Elsevier]
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- 2007
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6. Novel nucleotide triphosphates as potent P2Y2 agonists
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Brookings, Daniel, Davenport, Richard J., Davis, Jeremy, Galvin, Frances C.A., Lloyd, Steve, Mack, Stephen R., Owens, Ray, Sabin, Verity, and Wynn, Joanne
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NUCLEOTIDES , *PHOSPHATES , *ORGANIC compounds , *CELL receptors , *BIOSYNTHESIS - Abstract
Abstract: The synthesis and P2Y2 activities of a novel series of nucleoside triphosphates are described. Many of these compounds were potent agonists of the P2Y2 receptor. [Copyright &y& Elsevier]
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- 2007
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7. Benefits of Automated Crystallization Plate Tracking, Imaging, and Analysis
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Mayo, Christopher J., Diprose, Jonathan M., Walter, Thomas S., Berry, Ian M., Wilson, Julie, Owens, Ray J., Jones, E. Yvonne, Harlos, Karl, Stuart, David I., and Esnouf, Robert M.
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DATABASES , *CRYSTALLIZATION , *IMAGING systems , *SCANNING systems - Abstract
Summary: We describe the design of a database and software for managing and organizing protein crystallization data. We also outline the considerations behind the design of a fast web interface linking protein production data, crystallization images, and automated image analysis. The database and associated interfaces underpin the Oxford Protein Production Facility (OPPF) crystallization laboratory, collecting, in a routine and automatic manner, up to 100,000 images per day. Over 17 million separate images are currently held in this database. We discuss the substantial scientific benefits automated tracking, imaging, and analysis of crystallizations offers to the structural biologist: analysis of the time course of the trial and easy analysis of trials with related crystallization conditions. Features of this system address requirements common to many crystallographic laboratories that are currently setting up (semi-)automated crystallization imaging systems. [Copyright &y& Elsevier]
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- 2005
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8. The nsp9 Replicase Protein of SARS-Coronavirus, Structure and Functional Insights
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Sutton, Geoff, Fry, Elizabeth, Carter, Lester, Sainsbury, Sarah, Walter, Tom, Nettleship, Joanne, Berrow, Nick, Owens, Ray, Gilbert, Robert, Davidson, Andrew, Siddell, Stuart, Poon, Leo L.M., Diprose, Jonathan, Alderton, David, Walsh, Martin, Grimes, Jonathan M., and Stuart, David I.
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CORONAVIRUS diseases , *PROTEINS , *VIRAL replication , *RNA , *ULTRACENTRIFUGATION - Abstract
As part of a high-throughput structural analysis of SARS-coronavirus (SARS-CoV) proteins, we have solved the structure of the non-structural protein 9 (nsp9). This protein, encoded by ORF1a, has no designated function but is most likely involved with viral RNA synthesis. The protein comprises a single β-barrel with a fold previously unseen in single domain proteins. The fold superficially resembles an OB-fold with a C-terminal extension and is related to both of the two subdomains of the SARS-CoV 3C-like protease (which belongs to the serine protease superfamily). nsp9 has, presumably, evolved from a protease. The crystal structure suggests that the protein is dimeric. This is confirmed by analytical ultracentrifugation and dynamic light scattering. We show that nsp9 binds RNA and interacts with nsp8, activities that may be essential for its function(s). [Copyright &y& Elsevier]
- Published
- 2004
- Full Text
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