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Journal articleHaase S, Zimmermann D, Olshina MA, et al., 2015,
Disassembly activity of actin depolymerization factor (ADF) is associated with distinct cellular processes in apicomplexan parasites.
, Molecular Biology of the Cell, Vol: 26, Pages: 3001-3012, ISSN: 1939-4586Proteins of the actin depolymerizing factor (ADF)/cofilin family have been shown to be crucial for the motility and survival of apicomplexan parasites. However, the mechanisms by which ADF proteins fulfill their function remains poorly understood. In this study we sought to investigate the comparative activities of ADF proteins from Toxoplasma gondii and Plasmodium falciparum, the human malaria parasite, using a conditional T. gondii ADF-knockout line complemented with ADF variants from either species. We show that P. falciparum ADF1 can fully restore native TgADF activity, demonstrating functional conservation between parasites. Strikingly, mutation of a key basic residue (Lys(72)), previously implicated in disassembly in PfADF1, had no detectable phenotypic effect on parasite growth, motility or development. In contrast, organelle segregation was severely impaired when complementing with a TgADF mutant lacking the corresponding residue (Lys(68)). Biochemical analyses of each ADF protein confirmed the reduced ability of lysine mutants to mediate actin depolymerization via filament disassembly though not severing, in contrast to previous reports. These data suggest that actin filament disassembly is essential for apicomplexan parasite development but not for motility as well as pointing to genus-specific coevolution between ADF proteins and their native actin.
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Journal articleMcCarthy NL, Ces O, Law RV, et al., 2015,
Separation of liquid domains in model membranes induced with high hydrostatic pressure
, EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, Vol: 44, Pages: S115-S115, ISSN: 0175-7571 -
Journal articleMcCarthy NLC, Ces O, Law RV, et al., 2015,
Separation of liquid domains in model membranes induced with high hydrostatic pressure.
, Chem Commun (Camb), Vol: 51, Pages: 8675-8678We have imaged the formation of membrane microdomains immediately after their induction using a novel technology platform coupling high hydrostatic pressure to fluorescence microscopy. After formation, the ordered domains are small and highly dynamic. This will enhance links between model lipid assemblies and dynamic processes in cellular membranes.
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Journal articleElani Y, Law RV, Ces O, 2015,
Vesicle-based artificial cells: recent developments and prospects for drug delivery
, THERAPEUTIC DELIVERY, Vol: 6, Pages: 541-543, ISSN: 2041-5990- Author Web Link
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- Citations: 13
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Journal articleBranch T, Girvan P, Barahona M, et al., 2015,
Introduction of a Fluorescent Probe to Amyloid-β to Reveal Kinetic Insights into Its Interactions with Copper(II)
, ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, Vol: 54, Pages: 1227-1230, ISSN: 1433-7851- Author Web Link
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- Citations: 40
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Journal articleBarriga HMG, Booth P, Haylock S, et al., 2014,
Droplet interface bilayer reconstitution and activity measurement of the mechanosensitive channel of large conductance from Escherichia coli
, Journal of the Royal Society Interface, Vol: 11, ISSN: 1742-5662Droplet interface bilayers (DIBs) provide an exciting new platform for the study of membrane proteins in stable bilayers of controlled composition. To date, the successful reconstitution and activity measurement of membrane proteins in DIBs has relied on the use of the synthetic lipid 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC). We report the functional reconstitution of the mechanosensitive channel of large conductance (MscL) into DIBs composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), a lipid of significantly greater biological relevance than DPhPC. MscL functionality has been demonstrated using a fluorescence-based assay, showing that dye flow occurs across the DIB when MscL is gated by the cysteine reactive chemical 2-(trimethylammonium)ethyl methane thiosulfonate bromide (MTSET). MscL has already been the subject of a number of studies investigating its interaction with the membrane. We propose that this method will pave the way for future MscL studies looking in detail at the effects of controlled composition or membrane asymmetry on MscL activity using biologically relevant lipids and will also be applicable to other lipid–protein systems, paving the way for the study of membrane proteins in DIBs with biologically relevant lipids.
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Conference paperWalker RC, Dickson CJ, Madej BD, et al., 2014,
Amber lipid force field: Lipid14 and beyond
, 248th National Meeting of the American-Chemical-Society (ACS), Publisher: AMER CHEMICAL SOC, ISSN: 0065-7727 -
Journal articleDickson CJ, Madej BD, Skjevik AA, et al., 2014,
Lipid14: The Amber Lipid Force Field
, JOURNAL OF CHEMICAL THEORY AND COMPUTATION, Vol: 10, Pages: 865-879, ISSN: 1549-9618- Author Web Link
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- Citations: 868
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Conference paperNickdel MB, Lagarto JL, Kelly DJ, et al., 2014,
Autofluorescence lifetime metrology for label-free detection of cartilage matrix degradation
, Conference on Optical Biopsy XII, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X -
Conference paperMadej BD, Dickson CJ, Walker RC, et al., 2013,
Modular amber lipid force field for the simulation of complex membranes and membrane bound proteins
, 245th National Meeting of the American-Chemical-Society (ACS), Publisher: AMER CHEMICAL SOC, ISSN: 0065-7727
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