Paul Driscoll | Medical Research Council (original) (raw)

Papers by Paul Driscoll

The Journal of Organic Chemistry, 2003

Synthetic approaches to the lantibiotics, a family of thioether-bridged antimicrobial peptides, r... more Synthetic approaches to the lantibiotics, a family of thioether-bridged antimicrobial peptides, require flexible synthetic routes to a variety of orthogonally protected derivatives of lanthionine 1. The most direct approaches to lanthionine involve the reaction of cysteine with an alanyl beta-cation equivalent. Several possibilities exist for the alanyl beta-cation equivalent, including direct activation of serine under Mitsunobu conditions: however, the low reactivity of sulfur nucleophiles in the Mitsunobu reaction has previously precluded its use in the synthesis of the lantibiotics. We report here a new approach to the synthesis of protected lanthionine, using a novel variant of the Mitsunobu reaction in which catalytic zinc tartrate is used to enhance the nucleophilicity of the thiol. In the course of these studies, we have also demonstrated that the synthesis of lanthionine from trityl-protected beta-iodoalanines is prone to rearrangement, via an aziridine, to give predominantly trityl-protected alpha-iodo-beta-alanines, and hence norlanthionines, as the major products.

Page 364. 9 NMR of Nucleic Acids and Proteins BY PC DRISCOLL, D. ESPOSITO AND M. PFUHL 1 Introduc... more Page 364. 9 NMR of Nucleic Acids and Proteins BY PC DRISCOLL, D. ESPOSITO AND M. PFUHL 1 Introduction This is our fourth year of reporting on this very broad topic. As we have done in previous years we give the caveat ...

Journal of Biomolecular NMR, 2000

Lysozyme from lambda bacteriophage (k lysozyme) is an 18 kDa globular protein displaying some of ... more Lysozyme from lambda bacteriophage (k lysozyme) is an 18 kDa globular protein displaying some of the structural features common to all lysozymes; in particular, k lysozyme consists of two structural domains connected by a helix, and has its catalytic residues located at the interface between these two domains. An interesting feature of k lysozyme, when compared to the well-characterised hen egg-white lysozyme, is its lack of disulfide bridges; this makes k lysozyme an interesting system for studies of protein folding. A comparison of the folding properties of k lysozyme and hen lysozyme will provide important insights into the role that disulfide bonds play in the refolding pathway of the latter protein. Here we report the 1 H, 13 C and 15 N backbone resonance assignments for k lysozyme by heteronuclear multidimensional NMR spectroscopy. These assignments provide the starting point for detailed investigation of the refolding pathway using pulse-labelling hydrogen/deuterium exchange experiments monitored by NMR.

Journal of Biomolecular NMR, 2004

Annual Review of Biochemistry, 2001

The 3-phosphorylated inositol lipids fulfill roles as second messengers by interacting with the l... more The 3-phosphorylated inositol lipids fulfill roles as second messengers by interacting with the lipid binding domains of a variety of cellular proteins. Such interactions can affect the subcellular localization and aggregation of target proteins, and through allosteric effects, their activity. Generation of 3-phosphoinositides has been documented to influence diverse cellular pathways and hence alter a spectrum of fundamental cellular activities. This review is focused on the 3-phosphoinositide lipids, the synthesis of which is acutely triggered by extracellular stimuli, the enzymes responsible for their synthesis and metabolism, and their cell biological roles. Much knowledge has recently been gained through structural insights into the lipid kinases, their interaction with inhibitors, and the way their 3-phosphoinositide products interact with protein targets. This field is now moving toward a genetic dissection of 3-phosphoinositide action in a variety of model organisms. Such approaches will reveal the true role of the 3-phosphoinositides at the organismal level in health and disease.

Febs Letters, 2002

The FYVE zinc finger domain is conserved from yeast (five proteins) to man (27 proteins). It func... more The FYVE zinc finger domain is conserved from yeast (five proteins) to man (27 proteins). It functions in the membrane recruitment of cytosolic proteins by binding to phosphatidylinositol 3-phosphate (PI3P), which is found mainly on endosomes. Here we review recent work that sheds light on the targeting of FYVE finger proteins to PI3P-containing membranes, and how these proteins serve to

Journal of Experimental Medicine

The adhesion interaction between the immunoglobulin superfamily molecules CD2 and CD58 (lymphocyt... more The adhesion interaction between the immunoglobulin superfamily molecules CD2 and CD58 (lymphocyte function-associated antigen 3) plays an important role in T cell and natural killer cell interaction with various antigen-presenting and target cells. Determination of the solution structure of rat CD2 domain 1 has allowed a model of human CD2 domain 1 to be generated, and a series of mutants based on this model have been made. Residues of domain 1 of human CD2 predicted to be solvent exposed were substituted with the equivalent residues present in the rat CD2 molecule. The ability of these mutants to mediate rosetting with human and sheep erythrocytes was studied. Results show that the binding site of CD2 for both human and sheep CD58 maps to the beta sheet containing beta strands CC'C"F and G. Residues K34 and E36 in beta strand C, R48 and K49 in beta strand C', and K91 and N92 in the loop connecting beta strands F and G are shown to be critical in the interaction. The d...

Biochemistry

Modules which share the same consensus sequence are assumed to have common structural features, a... more Modules which share the same consensus sequence are assumed to have common structural features, at the secondary and tertiary level. In order to test the extent of such similarities, it is necessary to examine the structures of several examples from each module family. Recently, the first three-dimensional structure of a complement control protein (CCP) module (the 16th repeat of human factor H, H16) was determined using a combination of two-dimensional NMR and simulated annealing [Norman, D.G., Barlow, P.N., Baron, M., Day, A.J., Sim, R.B., & Campbell, I.D. (1991) J. Mol. Biol. 219, 717-725]. Using the same techniques, the three-dimensional structure of a second CCP module (the 5th repeat of human factor H, H5) has now been determined. The primary sequence of H5 contains 17 residues which are identical and in equivalent position to those in H16. Thirteen of these 17 are part of the consensus sequence. The similarities between the secondary structure of H5 and that of H16 are extens...

Methods in enzymology, 2014

This chapter describes reports of the structural characterization of death ligands and death rece... more This chapter describes reports of the structural characterization of death ligands and death receptors (DRs) from the tumor necrosis factor (TNF) and TNF receptor families. The review discusses the interactions of these proteins with agonist ligands, inhibitors, and downstream signaling molecules. Though historically labeled as being implicated in programmed cell death, the function of these proteins extends to nonapoptotic pathways. The review highlights, from a structural biology perspective, the complexity of DR signaling and the ongoing challenge to discern the precise mechanisms that occur at the point of DR activation, including how the degree to which the receptors are induced to cluster may be related to the nature of the impact upon the cell. The potential for posttranslational modification and receptor internalization to play roles in DR signaling is briefly discussed.

Biochemical Society transactions, 1993

Abbreviations used: LFA-3, lymphocyte function-associated antigen-3; RMSD, root-mean-square diffe... more Abbreviations used: LFA-3, lymphocyte function-associated antigen-3; RMSD, root-mean-square difference; IgSF, immunoglobulin superfamily; rCD2. D1, rat CD2 domain 1; NOESY, nuclear Overhauser effect spectroscopy; HOHAHA, homonuclear Hartmann-Hahn; HSQC, ...

Future Medicinal Chemistry, 2011

Successful structural investigations of protein-protein interactions can be facilitated by studyi... more Successful structural investigations of protein-protein interactions can be facilitated by studying only the core interacting regions of the constituent proteins. However, attempting the discovery of stable core complexes using informed trial-and-error approaches can prove time and resource intensive. We describe a valuable extension of combinatorial domain hunting (CDH), a technology for the timely elucidation of soluble protein truncations. The new method, CDH(2), enables empirical discovery of stable protein-protein core complexes. CDH(2) is demonstrated ab initio using a previously well-characterized Hsp90/Cdc37 complex. Without using a priori information, we demonstrate the isolation of stable protein-protein complexes, suitable for further analyses. This resource-efficient process can provide protein complexes for screening of compounds designed to modulate protein-protein interactions, thus facilitating novel drug discovery.

PLoS ONE

Death-associated protein kinase (DAPk) is a calcium/calmodulin-regulated Ser/Thr-protein kinase t... more Death-associated protein kinase (DAPk) is a calcium/calmodulin-regulated Ser/Thr-protein kinase that functions at an important point of integration for cell death signaling pathways. DAPk has a structurally unique multi-domain architecture, including a C-terminally positioned death domain (DD) that is a positive regulator of DAPk activity. In this study, recombinant DAPk-DD was observed to aggregate readily and could not be prepared in sufficient yield for structural analysis. However, DAPk-DD could be obtained as a soluble protein in the form of a translational fusion protein with the B1 domain of streptococcal protein G. In contrast to other DDs that adopt the canonical six amphipathic α-helices arranged in a compact fold, the DAPk-DD was found to possess surprisingly low regular secondary structure content and an absence of a stable globular fold, as determined by circular dichroism (CD), NMR spectroscopy and a temperature-dependent fluorescence assay. Furthermore, we measured th...

European Journal of Biochemistry, 1994

... Timothy J. DUDGEON', Matthew J. BOTTOMLEY', Paul C. DRISCOLL', Martin J. HUMPH... more ... Timothy J. DUDGEON', Matthew J. BOTTOMLEY', Paul C. DRISCOLL', Martin J. HUMPHRIES', A. Paul MOULD2, George I. WINGFIELD' and John M ... Analy-sis of this data by the program CONTIN (Provencher and Gloeckner, 1981 ; Provencher, 1982) estimates that VCAM-dl ,2 ...

Proceedings of the National Academy of Sciences of the United States of America, Jan 30, 2007

Type IV secretion (T4S) systems translocate DNA and protein effectors through the double membrane... more Type IV secretion (T4S) systems translocate DNA and protein effectors through the double membrane of Gram-negative bacteria. The paradigmatic T4S system in Agrobacterium tumefaciens is assembled from 11 VirB subunits and VirD4. Two subunits, VirB9 and VirB7, form an important stabilizing complex in the outer membrane. We describe here the NMR structure of a complex between the C-terminal domain of the VirB9 homolog TraO (TraO(CT)), bound to VirB7-like TraN from plasmid pKM101. TraO(CT) forms a beta-sandwich around which TraN winds. Structure-based mutations in VirB7 and VirB9 of A. tumefaciens show that the heterodimer interface is conserved. Opposite this interface, the TraO structure shows a protruding three-stranded beta-appendage, and here, we supply evidence that the corresponding region of VirB9 of A. tumefaciens inserts in the membrane and protrudes extracellularly. This complex structure elucidates the molecular basis for the interaction between two essential components of a...

Biochemistry, 1992

Page 1. 2068 Biochemistry 1992, 31, 2068-2073 spectively. This observation suggests interactions ... more Page 1. 2068 Biochemistry 1992, 31, 2068-2073 spectively. This observation suggests interactions of the re-spective vanadyl carrier complexes with biological recognition sites with more effective release of vanadyl ions from the L-enantiomeric complexes. ...

CD5 is a type-I transmembrane glycoprotein found on thymocytes, T-cells and a subset of B-cells. ... more CD5 is a type-I transmembrane glycoprotein found on thymocytes, T-cells and a subset of B-cells. The extracellular region consists of three domains belonging to the scavenger receptor cysteine-rich (SRCR) superfamily, for which no three-dimensional structure has been obtained. Recombinant soluble CD5 domain 1 (CD5d1), the N-terminal SRCR domain, has been expressed in both chinese hamster ovary (CHO) cells and Pichia pastoris. CD5d1 was shown to be correctly folded by binding to the CD5 monoclonal antibody Leu1. Circular dichroism and NMR analyses indicate that CD5d1 has a high β-sheet content. CD5d1 from both CHO cells and P. pastoris have very similar properties. The disulphide bonding pattern was determined and is consistent with that found for the group-A SRCR domain of type-1 macrophage scavenger receptor and MARCO, the macrophage receptor with collagenous structure. Observations have been made of the role of glycosylation of CD5. P. pastoris expression provides large quantities of correctly folded recombinant CD5d1 for multidimensional NMR and for X-ray crystallographic studies. The whole extracellular region of CD5, expressed as a chimaera with rat CD4 domains 3 and 4 (cCD5d1Ϫ3-CD4d3ϩ4), was studied by electron microscopy and carbohydrate analysis to gain an overview of the structure of the extracellular portion of intact CD5. Carbohydrate analysis identified N-linked glycans on CD5 domains 1 and 2, and sialylated O-linked glycans on the linker peptide between domains 1 and 2. Electron microscopy and carbohydrate analysis together suggest that the extracellular region of CD5 forms a rod-like structure with domain 1 distal from the cell surface and separated from domains 2 and 3 by an O-glycosylated peptide linker region.

Biomolecular NMR Assignments, 2008

Trends in Biochemical Sciences, 2002

Phosphoinositide 3-kinases (PI3Ks) are implicated in a variety of fundamental cellular processes.... more Phosphoinositide 3-kinases (PI3Ks) are implicated in a variety of fundamental cellular processes. These enzymes catalyse phosphorylation of the 3&amp;amp;amp;amp;amp;amp;amp;#39;-OH position of myo-inositol lipids that serve as secondary messengers. The catalytic subunit for one of the family members, PI3K gamma, has been structurally characterized, independently, in complexes with kinase inhibitors and with the p21(Ras) GTPase. These atomic structures provide a basis for the rationalization of some PI3K substrate specificities and regulatory mechanisms, establishing links to functional and cellular data. Ongoing comprehensive structural and functional studies are essential to realize the promise of PI3K isozyme-specific therapeutic agents.