呻吟之王

Dr Sarah Elizabeth Lee

Job: Researcher

Faculty: Health and Life Sciences

School/department: Leicester School of Pharmacy

Research group(s): Pharmacology

Address: 呻吟之王, The Gateway, Leicester, LE1 9BH

T: +44 (0)116 257 7954

E: s.lee@dmu.ac.uk

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Personal profile

Lee holds a degree in chemistry from Imperial College, London and a PhD from the University of Bangor looking at the mechanism and application of the non-classical Wittig reaction with carbohydrate-derived lactones. More recently, her research has strived to understand the role of phosphorus-containing chemicals in natural and unnatural biological processes. Specific research projects have included incorporating unnatural functionalities into nucleic acids, and the role of unnatural ATPs in elucidating cell signaling processes. Her research sits within the discipline of chemical biology, although her personal research looks at the issues presented in the systematic synthesis of the necessary chemical tools to probe the biological system.

Research group affiliations

Publications and outputs

dc.title: Exploring the roles of protein kinases using chemical genetics. dc.contributor.author: Elphick, Lucy M.; Lee, Sarah E.; Anderson, Alexandra A.; Child, Emma S.; Bonnac, Laurent; Gouverneur, Veronique; Mann, David J. dc.description.abstract: The protein kinase superfamily is one of the most important families of enzymes in molecular biology. Protein kinases typically catalyze the transfer of the 纬-phosphate from ATP to a protein substrate (a highly ubiquitous cellular reaction), thereby controlling key areas of cell regulation. Deregulation of protein kinases is known to contribute to many human diseases, and selective inhibitors of protein kinases are a major area of interest in medicinal chemistry. However, a detailed understanding of many kinase pathways is currently lacking. Before we can effectively design medicinally relevant selective kinase inhibitors, it is necessary to understand the role played by a given kinase in specific signal-transduction cascades and to decipher its protein targets. Here, we describe recent advances towards dissecting protein kinase function through the use of chemical genetics.
dc.title: Novel syntheses of (Z)-alkene and alkane base-modified nucleosides. dc.contributor.author: Lee, Sarah E.; Vyle, Joseph S.; Williams, David M.; Grasby, Jane A. dc.description.abstract: The syntheses of 5-(Z)-(3-aminoallyl)- and 5-(3-aminopropyl)-substituted 2鈥�-deoxyuridine and 2鈥�-deoxycytidine are reported. These compounds were derived from the corresponding 5-propargylamine derivative. [Hobbs, F. W. J. Org. Chem.1989, 52, 3420.] The catalyst we have employed for these reductions is a NiCl2/NaBH4 system, which we have found to be superior to the more conventional palladium-catalysts previously reported with similar compounds.
dc.title: Homometathesis and cross-metathesis coupling of phosphine-borane templates with electron-rich and electron-poor olefins. dc.contributor.author: Dunne, Katherine S.; Lee, Sarah E.; Gouverneur, Veronique dc.description.abstract: Ruthenium-catalysed olefin cross-metathesis can be used to synthesise structurally diverse acyclic phosphines protected as their borane complexes. Homodimerisations have been investigated and proved successful only for the allyl-substituted borane-protected phosphines. In the presence of various olefinic partners, allyl-substituted P templates reacted in cross-couplings to give predominantly the E products but traces of the Z isomers were always detected in the crude reaction mixtures. In contrast, cross-metathesis of vinyl-substituted phosphine boranes took place with exclusive E-selectivity. Although the conversions were consistently very good to excellent, the yields of purified products were often significantly lower suggesting that some of the newly formed compounds are prone to decompose upon purification.
dc.title: Using chemical genetics and ATP analogues to dissect protein kinase function. dc.contributor.author: Elphick, Lucy M.; Lee, Sarah E.; Gouverneur, Veronique; Mann, David J. dc.description.abstract: Protein kinases catalyze the transfer of the 纬-phosphate of ATP to a protein substrate and thereby profoundly alter the properties of the phosphorylated protein. The identification of the substrates of protein kinases has proven to be a very difficult task because of the multitude of structurally related protein kinases present in cells, their apparent redundancy of function, and the lack of absolute specificity of small-molecule inhibitors. Here, we review approaches that utilize chemical genetics to determine the functions and substrates of protein kinases, focusing on the design of ATP analogues and protein kinase binding site mutants.
dc.title: Enhancing the catalytic repertoire of nucleic acids: a systematic study of linker length and rigidity. dc.contributor.author: Lee, Sarah E.; Sidorov, Alexander; Gourlain, Thierry; Mignet, Nathalie; Thorpe, Simon J.; Brazier, John A.; Dickman, Mark J.; Hornby, David P.; Grasby, Jane A.; Williams, David M. dc.description.abstract: The incorporation of potentially catalytic groups in DNA is of interest for the in vitro selection of novel deoxyribozymes. A series of 10 C5-modified analogues of 2鈥�-deoxyuridine triphosphate have been synthesised that possess side chains of differing flexibility and bearing a primary amino or imidazole functionality. For each series of nucleotide analogues differing degrees of flexibility of the C5 side chain was achieved through the use of alkynyl, alkenyl and alkyl moieties. The imidazole function was conjugated to these C5-amino-modified nucleotides using either imidazole 4-acetic acid or imidazole 4-acrylic acid (urocanic acid). The substrate properties of the nucleotides (fully replacing dTTP) with Taq polymerase during PCR have been investigated in order to evaluate their potential applications for in vitro selection experiments. 5-(3-Aminopropynyl)dUTP and 5-(E-3-aminopropenyl)dUTP and their imidazole 4-acetic acid- and urocanic acid-modified conjugates were found to be substrates. In contrast, C5-amino-modified dUTPs with alkane or Z-alkene linkers and their corresponding conjugates were not substrates. The incorporation of these analogues during PCR has been confirmed by inhibition of restriction enzyme digestion using XbaI and by mass spectrometry of the PCR products.
dc.title: The synthesis of modified 5-(Aminoalkyl)- and 5-(Aminoalkenyl)- Uridine 5'-Triphosphates. dc.contributor.author: Lee, Sarah E.; Mignet, Nathalie; Vyle, Joseph S.; Whetton, L.; Grasby, Jane A.; Williams, David M.
dc.title: Enhancing the catalytic repertoire of nucleic acids. II. Simultaneous incorporation of amino and imidazolyl functionalities by two modified triphosphates during PCR. dc.contributor.author: Gourlain, Thierry; Sidorov, Alexander; Mignet, Nathalie; Thorpe, Simon J.; Lee, Sarah E.; Grasby, Jane A.; Williams, David M. dc.description.abstract: The incorporation of potentially catalytic groups into DNA is of interest for the in vitro selection of novel deoxyribozymes. We have devised synthetic routes to a series of three C7 modified 7-deaza-dATP derivatives with pendant aminopropyl, Z-aminopropenyl and aminopropynyl side chains. These modified triphosphates have been tested as substrates for Taq polymerase during PCR. All the modifications are tolerated by this enzyme, with the aminopropynyl side chain giving the best result. Most protein enzymes have more than one type of catalytic group located in their active site. By using C5-imidazolyl-modified dUTPs together with 3-(aminopropynyl)-7-deaza-dATP in place of the natural nucleotides dTTP and dATP, we have demonstrated the simultaneous incorporation of both amino and imidazolyl moieties into a DNA molecule during PCR. The PCR product containing the four natural bases was fully digested by XbaI, while PCR products containing the modified 7-deaza-dATP analogues were not cleaved. Direct evidence for the simultaneous incorporation during PCR of an imidazole-modified dUTP and an amino-modified 7-deaza-dATP has been obtained using mass spectrometry.
dc.title: Recent synthetic applications of the non-classical Wittig reaction. dc.contributor.author: Murphy, Patrick J.; Lee, Sarah E.
dc.title: A Quantitative Comparison of Wild-Type and Gatekeeper Mutant Cdk2 for Chemical Genetic Studies with ATP Analogues dc.contributor.author: Elphick, Lucy M.; Lee, Sarah E.; Child, Emma S.; Prasad, Aarathi; Pignocchi, Cristina; Thibaudeau, S茅bastien; Anderson, Alexandra A.; Bonnac, Laurent; Gouverneur, Veronique; Mann, David J.
dc.title: Synthesis and reactivity of novel c-phosphate modified ATP analogues dc.contributor.author: Lee, Sarah E.; Elphick, Lucy M.; Anderson, Alexandra A.; Bonnac, Laurent; Child, Emma S.; Mann, David J.

Research interests/expertise

Chemical synthesis (organic)
Modified carbohydrates
Modified ATPs
Organo-phosphorus chemistry
Chemistry of Phosphates
HPLC
Ion-exchange chromatography
Nucleosides, Nucleotides and Nucleic Acids