Protein surface functionalisation as a general strategy for facilitating biomimetic mineralisation of ZIF-8

The durability of enzymes in harsh conditions can be enhanced by encapsulation within metal-organic frameworks (MOFs) via a process called biomimetic mineralisation. Herein we show that the surface charge and chemistry of a protein determines its ability to seed MOF growth. We demonstrate that chemical modification of amino acids on the protein surface is an effective method for systematically controlling biomimetic mineralisation by zeolitic imidazolate framework-8 (ZIF-8). Reaction of surface lysine residues with succinic (or acetic) anhydride facilitates biomimetic mineralisation by increasing the surface negative charge, whereas reaction of surface carboxylate moieties with ethylenediamine affords a more positively charged protein and hinders the process. Moreover, computational studies confirm that the surface electrostatic potential of a protein is a good indicator of its ability to induce biomimetic mineralisation. This study highlights the important role played by protein surface chemistry in encapsulation and outlines a general method for facilitating the biomimetic mineralisation of proteins

Fused pyrazino [2,3-b]indolizine and indolizino[2,3-b]quioxaline derivatives; synthesis, structures and properties

The synthesis of six new compounds incorporating either a pyrazino[2,3-b]indolizine or indolizino[2,3-b]quinoxaline core are reported in good yield (58-87%). The intermediates for the key cyclization reaction for one set of compounds (5a-c), with a sterically demanding 3,5-dimethylpyrazole group in the 5-position of the core, were found to be mono-substituted. These intermediates could be isolated and cyclized by heating under acid-catalyzed conditions. To further demonstrate the versatility of the chemistry, compounds 6a-c were synthesized in 58-68% yields. Compounds 5a-c are non-planar in solution and the solid-state, while 6a-c have close to planar conformations, pointing to weak hydrogen bonds between the acidic C-Hs and the adjacent azine nitrogen atoms. The cytotoxicity of the six newly synthesized and three previously prepared compounds was assessed against a human glioblastoma multiforme cell line. © 2011 Elsevier Ltd. All rights reserved.Witold M. Bloch, Stephanie M. Derwent-Smith, Fatiah Issa, Jonathan C. Morris, Louis M. Rendina and Christopher J. Sumbyhttp://www.journals.elsevier.com/tetrahedron

Some reactions of azides with diynyl-bis(phosphine)ruthenium-cyclopentadienyl complexes

Abstract not availableMichael I. Bruce, Alexandre Burgun, Jonathan George, Brian K. Nicholson, Christian R. Parker, Brian W. Skelton, Nancy Scoleri, Christopher J. Sumby, Natasha N. Zaitsev

Structural systematics of some trinuclear alkynyl and diynyl Group 11 complexes containing dppm [dppm = CH(2)(PPh(2))(2)]

Available online 9 October 2017In this review the molecular structures of a series of trinuclear alkynyl and diynyl Group 11 cations [{M3(μ-dppm)3}(X)n](3−n)+ (M = Cu, Ag; n = 1, 2; where X is an alkynyl or diynyl group, an inorganic anion or solvent) are considered from the points of view of (i) the dimensions and geometries of the M3(P–P)3 cores, (ii) the conformations of the dppm ligands, and (iii) the attachment of the alkynyl and diynyl ligands. In the crowded [M3(μ-dppm)3]3+ core, the dppm ligands are arranged so that there is always one CH2 group up and two down, to give pseudo mirror symmetry perpendicular to the M3 plane (crystallographic in some cases). Attachment of the alkynyl or diynyl substituent(s) occurs roughly normal to the M3 plane; according to their perpendicularity, the C(1) atom may be μ2 or μ3. In most cases where only one alkynyl or diynyl ligand is present, a second ligand is also attached to the M3 core. Unusual and interesting dispositions/conformations of the dppm ligands are widespread, among the mono–diynyl complexes in particular, whereby some phosphorus donor atoms lie at unusual distances out of the M3 planes, a concomitant of strong agostic interactions between phenyl H atoms and the atoms of the open M3 face, and weak M⋯M interactions. With one X group, C–H⋯M interactions persist on the other face, with C–H⋯X interactions with the alkyne affecting the inclination of the alkyne and the conformation of the Ph rings. With two substituents (one of which may be a loosely bound anion), similar interactions may occur, accompanied by twisting of the dppm chelate ring to displace P atoms from the M3 plane. These factors possibly inhibit formation of the bis(diyndiyl) complexes, which are only obtained under more strongly basic conditions.Michael I. Bruce, Jean-François Halet, Boris Le Guennic, Brian W. Skelton, Alexandre N. Sobolev, Christopher J. Sumby, Allan H. Whit

Incompetence of Neutrophils to Invasive Group A streptococcus Is Attributed to Induction of Plural Virulence Factors by Dysfunction of a Regulator

Group A streptococcus (GAS) causes variety of diseases ranging from common pharyngitis to life-threatening severe invasive diseases, including necrotizing fasciitis and streptococcal toxic shock-like syndrome. The characteristic of invasive GAS infections has been thought to attribute to genetic changes in bacteria, however, no clear evidence has shown due to lack of an intriguingly study using serotype-matched isolates from clinical severe invasive GAS infections. In addition, rare outbreaks of invasive infections and their distinctive pathology in which infectious foci without neutrophil infiltration hypothesized us invasive GAS could evade host defense, especially neutrophil functions. Herein we report that a panel of serotype-matched GAS, which were clinically isolated from severe invasive but not from non-invaive infections, could abrogate functions of human polymorphnuclear neutrophils (PMN) in at least two independent ways; due to inducing necrosis to PMN by enhanced production of a pore-forming toxin streptolysin O (SLO) and due to impairment of PMN migration via digesting interleukin-8, a PMN attracting chemokine, by increased production of a serine protease ScpC. Expression of genes was upregulated by a loss of repressive function with the mutation of csrS gene in the all emm49 severe invasive GAS isolates. The csrS mutants from clinical severe invasive GAS isolates exhibited high mortality and disseminated infection with paucity of neutrophils, a characteristic pathology seen in human invasive GAS infection, in a mouse model. However, GAS which lack either SLO or ScpC exhibit much less mortality than the csrS-mutated parent invasive GAS isolate to the infected mice. These results suggest that the abilities of GAS to abrogate PMN functions can determine the onset and severity of invasive GAS infection

Unveiling the structural transitions during activation of a CO2 methanation catalyst Ru0/ZrO2 synthesised from a MOF precursor

Available online 5 May 2020Carbon Capture, Utilisation and Storage (CCUS) technologies are utilised to minimise net CO2 emissions and hence mitigate the impact of anthropogenic emissions on the global climate. One example of CO2 utilisation is the production of carbon-neutral methane fuel via catalytic CO2 reduction with H2 (methanation). Thermal activation of a metal impregnated metal-organic framework (MOF), 1 wt%Ru/UiO-66 in the presence of H2 and CO2 provides in situ synthesis of a highly active methanation catalyst: H2 promotes the formation of Ru0 nanoparticles, and CO2 behaves as a mild oxidant to remove framework carbon and promote ZrO2 crystallisation. The nature of the active MOF-derived Ru0/ZrO2 catalyst was studied by PXRD, TEM, and XAS, and the evolution of the parent 1 wt%Ru/UiO-66 during thermal activation monitored in operando by synchrotron PXRD. The Ru impregnated Zr-based MOF collapses on heating in H2 and CO2 to form an amorphous C and Zr containing phase that subsequently crystallises as tetragonal (t-) ZrO2 nanoparticles. These t-ZrO2 nanoparticles undergo a subsequent phase transition to the more stable monoclinic (m-) ZrO2 polymorph. In situ activation of Ru/UiO-66 generates a highly active catalyst for CO2 methanation by transforming the MOF precursor into a (carbonfree) crystalline t-ZrO2 support that stabilises highly dispersed metallic Ru nanoparticles. This insight may guide the rational design of future MOF-derived catalystsRenata Lippi, Anita M. D, Angelo, Chaoen Li, Shaun C. Howard, Ian C. Madsen, Karen Wilson, Adam F. Lee, Christopher J. Sumby, Christian J. Doonan, Jim Patel, Danielle F. Kenned

Allelic replacement of the streptococcal cysteine protease SpeB in a Δsrv mutant background restores biofilm formation

<p>Abstract</p> <p>Background</p> <p>Group A <it>Streptococcus </it>(GAS) is a Gram-positive human pathogen that is capable of causing a wide spectrum of human disease. Thus, the organism has evolved to colonize a number of physiologically distinct host sites. One such mechanism to aid colonization is the formation of a biofilm. We have recently shown that inactivation of the streptococcal regulator of virulence (Srv), results in a mutant strain exhibiting a significant reduction in biofilm formation. Unlike the parental strain (MGAS5005), the streptococcal cysteine protease (SpeB) is constitutively produced by the <it>srv </it>mutant (MGAS5005Δ<it>srv</it>) suggesting Srv contributes to the control of SpeB production. Given that SpeB is a potent protease, we hypothesized that the biofilm deficient phenotype of the <it>srv </it>mutant was due to the constitutive production of SpeB. In support of this hypothesis, we have previously demonstrated that treating cultures with E64, a commercially available chemical inhibitor of cysteine proteases, restored the ability of MGAS5005Δ<it>srv </it>to form biofilms. Still, it was unclear if the loss of biofilm formation by MGAS5005Δ<it>srv </it>was due only to the constitutive production of SpeB or to other changes inherent in the <it>srv </it>mutant strain. To address this question, we constructed a Δ<it>srv</it>Δ<it>speB </it>double mutant through allelic replacement (MGAS5005Δ<it>srv</it>Δ<it>speB</it>) and tested its ability to form biofilms <it>in vitro</it>.</p> <p>Findings</p> <p>Allelic replacement of <it>speB </it>in the <it>srv </it>mutant background restored the ability of this strain to form biofilms under static and continuous flow conditions. Furthermore, addition of purified SpeB to actively growing wild-type cultures significantly inhibited biofilm formation.</p> <p>Conclusions</p> <p>The constitutive production of SpeB by the <it>srv </it>mutant strain is responsible for the significant reduction of biofilm formation previously observed. The double mutant supports a model by which Srv contributes to biofilm formation and/or dispersal through regulation of <it>speB</it>/SpeB.</p

Group A Streptococcus Secreted Esterase Hydrolyzes Platelet-Activating Factor to Impede Neutrophil Recruitment and Facilitate Innate Immune Evasion

The innate immune system is the first line of host defense against invading organisms. Thus, pathogens have developed virulence mechanisms to evade the innate immune system. Here, we report a novel means for inhibition of neutrophil recruitment by Group A Streptococcus (GAS). Deletion of the secreted esterase gene (designated sse) in M1T1 GAS strains with (MGAS5005) and without (MGAS2221) a null covS mutation enhances neutrophil ingress to infection sites in the skin of mice. In trans expression of SsE in MGAS2221 reduces neutrophil recruitment and enhances skin invasion. The sse deletion mutant of MGAS5005 (ΔsseMGAS5005) is more efficiently cleared from skin than the parent strain. SsE hydrolyzes the sn-2 ester bond of platelet-activating factor (PAF), converting biologically active PAF into inactive lyso-PAF. KM and kcat of SsE for hydrolysis of 2-thio-PAF were similar to those of the human plasma PAF acetylhydrolase. Treatment of PAF with SsE abolishes the capacity of PAF to induce activation and chemotaxis of human neutrophils. More importantly, PAF receptor-deficient mice significantly reduce neutrophil infiltration to the site of ΔsseMGAS5005 infection. These findings identify the first secreted PAF acetylhydrolase of bacterial pathogens and support a novel GAS evasion mechanism that reduces phagocyte recruitment to sites of infection by inactivating PAF, providing a new paradigm for bacterial evasion of neutrophil responses

Dispersal of Group A Streptococcal Biofilms by the Cysteine Protease SpeB Leads to Increased Disease Severity in a Murine Model

Group A Streptococcus (GAS) is a Gram-positive human pathogen best known for causing pharyngeal and mild skin infections. However, in the 1980's there was an increase in severe GAS infections including cellulitis and deeper tissue infections like necrotizing fasciitis. Particularly striking about this elevation in the incidence of severe disease was that those most often affected were previously healthy individuals. Several groups have shown that changes in gene content or regulation, as with proteases, may contribute to severe disease; yet strains harboring these proteases continue to cause mild disease as well. We and others have shown that group A streptococci (MGAS5005) reside within biofilms both in vitro and in vivo. That is to say that the organism colonizes a host surface and forms a 3-dimensional community encased in a protective matrix of extracellular protein, DNA and polysaccharide(s). However, the mechanism of assembly or dispersal of these structures is unclear, as is the relationship of these structures to disease outcome. Recently we reported that allelic replacement of the streptococcal regulator srv resulted in constitutive production of the streptococcal cysteine protease SpeB. We further showed that the constitutive production of SpeB significantly decreased MGAS5005Δsrv biofilm formation in vitro. Here we show that mice infected with MGAS5005Δsrv had significantly larger lesion development than wild-type infected animals. Histopathology, Gram-staining and immunofluorescence link the increased lesion development with lack of disease containment, lack of biofilm formation, and readily detectable levels of SpeB in the tissue. Treatment of MGAS5005Δsrv infected lesions with a chemical inhibitor of SpeB significantly reduced lesion formation and disease spread to wild-type levels. Furthermore, inactivation of speB in the MGAS5005Δsrv background reduced lesion formation to wild-type levels. Taken together, these data suggest a mechanism by which GAS disease may transition from mild to severe through the Srv mediated dispersal of GAS biofilms