BSc Chemistry, University of Bristol (1991-1994); PhD Chemistry, University of Bristol (1994-1997) Post-doctoral Research Fellow, SmithKline Beecham (1998-2000); World Travel (2000-2001); Experimental Officer, University of Southampton (2001-).

Southampton Biomolecular NMR
Biomolecular NMR at Southampton revolves around protein structure, dynamics and interactions although we also carry out small molecule work on biofluids and enzyme products. Most of the data is acquired on our 600MHz Varian spectrometer although we have use of the 800 and 900MHz facilities at Mill Hill and Birmingham.

1. Calmodulin binding of glutamate receptor peptides

Over-activation of metabotropic glutamate receptors (mGluR) is implicated in a variety of diseases such as epilepsy, Parkinson's and Huntington's diseases as well as in pain, schizophrenia and anxiety. mGluR7 regulates the glutamate levels in the synaptic cleft by providing feedback inhibition of the glutamate release mechanism.

The binding of calmodulin to a region on mGluR7 ensures the inhibition is maintained until glutamate concentrations have returned to a safe level. We're currently using residual dipolar coupling data and dynamics to structurally characterise the binding mode. Collaboration with Vincent O'Connor .

Figure 1. The 1-14 binding mode of calmodulin (red & green) to a peptide (blue). The 1-14 describes the spacing of 2 hydrophobic residues (shown in ball and stick) that are important in the interaction. The grey spheres are the 4 Ca 2+ ions.

For info on calmodulin see http://calcium.uhnres.utoronto.ca/ctdb/ctdb/home.html

2. HIV capsid domain binding of inhibitory peptide

Formation of infectious HIV-1 involves assembly of Gag polyproteins into immature particles and subsequent assembly of mature capsids after proteolytic disassembly of the Gag shell. A capsid assembly inhibitor (CAI), that binds the capsid domain of Gag and inhibits assembly of immature- and mature-like capsid particles in vitro has been identified by our collaborator Hans-Georg Kräusslich in Heidelberg . We have used chemical shift mapping to locate the binding site of the peptide on the capsid. http://news.bbc.co.uk/1/hi/health/4707449.stm - http://www.nature.com/nsmb/journal/vaop/ncurrent/abs/nsmb964.html

3. Protein L interactions

The bacterial surface protein, protein L, is a bacterial virulence factor, the expression of which is correlated with bacterial vaginosis and is thus of great interest currently. The interaction of protein L with itself and mammalian immunoglobulin light chains and the effects of mutations within protein L have been studied by NMR chemical shift mapping at Southampton . Collaboration with Mike Gore .

Evidence for plasticity and structural mimicry at the immunoglobulin light chain protein L interface. J Biol Chem. 2002, 277, 47500-47506.

4. Oxytetracycline acyl carrier protein structure

Certain bacteria possess the ability to manufacture useful therapeutics such as antibiotics. Streptomyces rimosus produces oxytetracycline which is commonly prescribed for the treatment of acne. This research has the principal aim of understanding the mechanisms by which such antibiotics are assembled in the hope of modifying the pathways to produce novel and more potent therapeutics. This research takes a structural approach to the problem and we have good links with Matt Crump in the polyketide group at Bristol .

Solution Structure and Dynamics of Oxytetracycline ACP from Streptomyces rimosus Biochemistry, 2003, 42, 8423-8433.

5. Cyclic DNA and Quadruplexes

In collaboration with Keith Fox and Tom Brown we have looked at DNA quadruplexes (Figure x) and miniature cyclic DNA duplexes of consisting only two basepairs. Identification of base-pairing is easy using NMR since the imino-protons are shifted to 11ppm or more (Figure y).

Figure x

Figure y

The structure of the cyclic DNA 2mer has been solved with the help of undergraduate Jasmin Baxter as her final year project and Dr John Stuart. Base-paired protons were barely detectable at 25C for this complex but were very prominent at lower temperatures. Figure z below shows the chemical structure, the imino proton region of the NMR spectrum at different temperatures and the ensemble of 3D structures derived from NOEs.

Further information on NMR projects can be found on our group website here.

A Scheschonka, S Findlow, R Schemm, O El Far, JH Caldwell, MP Crump, K Holden-Dye, V O'Connor, H Betz, JM Werner. Structural determinants of calmodulin binding to the intracellular C-terminal domain of the metabotropic glutamate receptor 7. A.J. Mol. Biol., 2008, 283, 5577-88.

AH El-Sagheer, R Kumar, S Findlow, JM Werner, AN Lane, T Brown. A very stable cyclic DNA miniduplex with just two base pairs. Chem. Biochem., 2008, 9, 50-52.

PA Rachwal, IS Findlow, JM Werner, T Brown and KR Fox. Intramolecular DNA quadruplexes with different arrangements of short and long loops. Nucleic Acids Research, 2007, 35, 4214-22.

Christopher J. Arthur, Anna E. Szafranska, Jed Long, Jane Mills, Russell J. Cox, Stuart C. Findlow, Thomas J. Simpson, Matthew P. Crump and John Crosby. The Malonyl Transferase Activity of Type II Polyketide Synthase Acyl Carrier Proteins
Chemistry & Biology, 2006, 13, 587-596

P. T. Erskine, G. D. E. Beaven, R. Hagan, I. S. Findlow, J. M. Werner S. P. Wood, J. Vernon, K. P. Giese, G. Fox and J. B. Cooper. Structure of the Neuronal Protein Calexcitin Suggests a Mode of Interaction in Signalling Pathways of Learning and Memory. J. Mol. Biol., 2006, 357, 1537-47.

Christopher J. Arthur, Anna Szafranska, Simon Evans, Stuart C. Findlow, Steve Burston, Ian Clark-Lewis, Thomas J. Simpson, John Crosby, Matthew P. Crump Self-malonylation is an intrinsic property of a chemical synthesized Type II polyketide synthase acyl carrier protein. Biochemistry, 2005, 44, 15414-15421.

Jana Sticht, Michael Humbert, Stuart Findlow, Jochen Bodem, Barbara Muller, Ursula Dietrich, Jorn Werner, Hans-Georg Krausslich. A Peptide Inhibitor of HIV-1 Assembly. Nature – Structural and Molecular Biology, 2005, 12, 671-677.

Matthew P. Crump, Thomas A. Ceska, Leo Spyracopoulos, Alistair Henry, Sarah C. Archibald, Rikki Alexander, Richard J. Taylor, Stuart C. Findlow, James O’Connell, Martyn K. Robinson and Anthony Shock. The structure of an allosteric inhibitor of LFA-1 bound to the I-domain studied by crystallography, NMR and calorimetry. Biochemistry, 2004, 43, 2394-404.

Stuart C. Findlow, Claire Winsor, Thomas J. Simpson, John Crosby, Matthew P. Crump. Solution Structure and Dynamics of Oxytetracycline Polyketide Synthase Acyl Carrier Protein from Streptomyces rimosus. Biochemistry, 2003, 42, 8423-8433

Graille M, Harrison S, Crump MP, Findlow SC, Housden NG, Muller BH, Battail-Poirot N, Sibai G, Sutton BJ, Taussig MJ, Jolivet-Reynaud C, Gore MG, Stura EA. Evidence for plasticity and structural mimicry at the immunoglobulin light chain protein L interface. J Biol Chem. 2002, 277, 47500-47506.

Hubbard JA, MacLachlan LK, Johnson P, Findlow SC, Ladds JC, Lewis CJ, Carter PS, Jones JJ, Fosberry AP. A method for identification of inhibitors of the phosphorylation reactions of bacterial response regulator proteins using (31)P nuclear magnetic resonance spectroscopy. Anal Biochem. 2001, 299, 31-6.

Crosby J, Byrom KJ, Hitchman TS, Cox RJ, Crump MP, Findlow IS, Bibb MJ, Simpson TJ. Acylation of Streptomyces type II polyketide synthase acyl carrier proteins. FEBS Lett. 1998, 433,132-8.

Cox RJ, Hitchman TS, Byrom KJ, Findlow IS, Tanner JA, Crosby J, Simpson TJ. Post-translational modification of heterologously expressed Streptomyces type II polyketide synthase acyl carrier proteins. FEBS Lett. 1997, 405, 267-72.

Findlow S, Gaskin P, Harrison PA, Lenton JR, Penny M, Willis CL. Syntheses of gibberellins A(93) and A(94) natural products detected in wheat grain. Journal Of The Chemical Society-Perkin Transactions 1, 1997, 5, 751-757.

Updated August 2008