Browsing by Author "Harrop, SJ"

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    CLIC proteins, ezrin, radixin, moesin and the coupling of membranes to the actin cytoskeleton: A smoking gun?
    (Elsevier, 2014-02-01) Jiang, L; Phang, JM; Yu, J; Harrop, SJ; Sokolova, AV; Duff, AP; Wilik, KE; Alkhamici, H; Breit, SN; Valenzuela, SM; Brown, LJ; Curmi, PMG
    The CLIC proteins are a highly conserved family of metazoan proteins with the unusual ability to adopt both soluble and integral membrane forms. The physiological functions of CLIC proteins may include enzymatic activity in the soluble form and anion channel activity in the integral membrane form. CLIC proteins are associated with the ERM proteins: ezrin, radixin and moesin. ERM proteins act as cross-linkers between membranes and the cortical actin cytoskeleton. Both CLIC and ERM proteins are controlled by Rho family small GTPases. CLIC proteins, ERM and Rho GTPases act in a concerted manner to control active membrane processes including the maintenance of microvillar structures, phagocytosis and vesicle trafficking. All of these processes involve the interaction of membranes with the underlying cortical actin cytoskeleton. The relationships between Rho GTPases, CLIC proteins, ERM proteins and the membrane:actin cytoskeleton interface are reviewed. Speculative models are proposed involving the formation of localised multi-protein complexes on the membrane surface that assemble via multiple weak interactions. This article is part of a Special Issue entitled: Reciprocal influences between cell cytoskeleton and membrane channels, receptors and transporters. © 2013, Elsevier B.V.
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    Crystal structure of posnjakite formed in the first crystal water-cooling line of the ANSTO Melbourne Australian Synchrotron MX1 Double Crystal Monochromator
    (International Union of Crystallography (IUCr), 2020-06-30T14:00:00Z) Mills, SJ; Aishima, J; Aragao, D; Caradoc-Davies, TT; Cowieson, NP; Gee, CL; Ericsson, D; Harrop, SJ; Panjikar, S; Smith, KML; Riboldi-Tunnicliffe, A; Williamson, R; Price, JR
    Exceptionally large crystals of posnjakite, CuSO(OH)(HO), formed during corrosion of a Swagelock(tm) Snubber copper gasket within the MX1 beamline at the ANSTO-Melbourne, Australian Synchrotron. The crystal structure was solved using synchrotron radiation to = 0.029 and revealed a structure based upon [Cu(OH)(HO)O] sheets, which contain Jahn-Teller-distorted Cu octa-hedra. The sulfate tetra-hedra are bonded to one side of the sheet corner sharing and linked to successive sheets extensive hydrogen bonds. The sulfate tetra-hedra are split and rotated, which enables additional hydrogen bonds. © Mills et al. 2020.
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    Selective inhibition of human group IIA-secreted phospholipase A(2) (hGIIA) signaling reveals arachidonic acid metabolism Is associated with colocalization of hGIIA to vimentin in rheumatoid synoviocytes
    (Americal Society Biochemistry Molecular Biology Inc., 2013-05-24) Lee, LK; Bryant, KJ; Bouveret, R; Lei, PW; Duff, AP; Harrop, SJ; Huang, EP; Harvey, RP; Gelb, MH; Gray, PP; Curmi, PMG; Cunningham, AM; Church, WB; Scott, KF
    Human group IIA secreted phospholipase A(2) (hGIIA) promotes tumor growth and inflammation and can act independently of its well described catalytic lipase activity via an alternative poorly understood signaling pathway. With six chemically diverse inhibitors we show that it is possible to selectively inhibit hGIIA signaling over catalysis, and x-ray crystal structures illustrate that signaling involves a pharmacologically distinct surface to the catalytic site. We demonstrate in rheumatoid fibroblast-like synoviocytes that non-catalytic signaling is associated with rapid internalization of the enzyme and colocalization with vimentin. Trafficking of exogenous hGIIA was monitored with immunofluorescence studies, which revealed that vimentin localization is disrupted by inhibitors of signaling that belong to a rare class of small molecule inhibitors that modulate protein-protein interactions. This study provides structural and pharmacological evidence for an association between vimentin, hGIIA, and arachidonic acid metabolism in synovial inflammation, avenues for selective interrogation of hGIIA signaling, and new strategies for therapeutic hGIIA inhibitor design. © 2013, American Society for Biochemistry and Molecular Biology
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    Structural characterization by small angle scattering suggests models for monomeric and dimeric forms of full-length ezrin
    (Australian Institute of Nuclear Science and Engineering, 2016-11-29) Phang, JM; Harrop, SJ; Duff, AP; Sokolova, AV; Crossett, B; Walsh, JC; Beckham, SA; Nguyen, CD; Davies, RB; Glöckner, C; Bromley, EH; Wilk, KE; Curmi, PM
    Ezrin is member of the ERM (Ezrin-Radixin-Moesin) family of proteins that have been conserved through metazoan evolution. These proteins have dormant and active forms, there the latter links the actin cytoskeleton to membranes. ERM proteins have three domains: an N-terminal FERM (band Four-point-one ERM) domain comprising three subdomains (F1, F2 and F3); a helical domain; and a C-terminal actin-binding domain. In the dormant form, FERM and C-terminal domains form a stable complex. We have determined crystal structures of the active FERM domain and the dormant FERM:C-terminal domain complex of human ezrin. We observe bistable array of phenylalanine residues in the core of subdomain F3 that is mobile in the active form and locked in the dormant form. As subdomain F3 is pivotal in binding membrane proteins and phospholipids, these transitions may facilitate activation and signaling. Full-length ezrin forms stable monomers and dimers. We used small-angle x-ray scattering to determine the solution structures of these species. As expected, the monomer shows a globular domain with a protruding helical coiled-coil. The dimer shows an elongated dumbbell structure that is twice as long as the monomer. By aligning ERM sequences spanning metazoan evolution, we show that the central helical region is conserved, preserving the heptad repeat. Using this, we have built a dimer model where each monomer forms half of an elongated anti-parallel coiled-coil with domain swapped FERM:C-terminal domain complexes at each end. The model suggests that ERM dimers may bind to actin in a parallel fashion.