Controlling Spin Crossover in a Family of Dinuclear Fe(III) Complexes via the Bis(catecholate) Bridging Ligand.

Spin crossover (SCO) complexes can reversibly switch between low spin (LS) and high spin (HS) states, affording possible applications in sensing, displays, and molecular electronics. Dinuclear SCO complexes with access to [LS-LS], [LS-HS], and [HS-HS] states may offer increased levels of functionality. The nature of the SCO interconversion in dinuclear complexes is influenced by the local electronic environment. We report the synthesis and characterization of [{FeIII(tpa)}2spiro](PF6)2 (1), [{FeIII(tpa)}2Br4spiro](PF6)2 (2), and [{FeIII(tpa)}2thea](PF6)2 (3) (tpa = tris(2-pyridylmethyl)amine, spiroH4 = 3,3,3',3'-tetramethyl-1,1'-spirobi(indan)-5,5',6,6'-tetraol, Br4spiroH4 = 3,3,3',3'-tetramethyl-1,1'-spirobi(indan)-4,4',7,7'-tetrabromo-5,5',6,6'-tetraol, theaH4 = 2,3,6,7-tetrahydroxy-9,10-dimethyl-9,10-dihydro-9,10-ethanoanthracene), utilizing non-conjugated bis(catecholate) bridging ligands. In the solid state, magnetic and structural analysis shows that 1 remains in the [HS-HS] state, while 2 and 3 undergo a partial SCO interconversion upon cooling from room temperature involving the mixed [LS-HS] state. In solution, all complexes undergo SCO from [HS-HS] at room temperature, via [LS-HS] to mixtures including [LS-LS] at 77 K, with the extent of SCO increasing in the order 1 < 2 < 3. Gas phase density functional theory calculations suggest a [LS-LS] ground state for all complexes, with the [LS-HS] and [HS-HS] states successively destabilized. The relative energy separations indicate that ligand field strength increases following spiro4- < Br4spiro4- < thea4-, consistent with solid-state magnetic and EPR behavior. All three complexes show stabilization of the [LS-HS] state in relation to the midpoint energy between [LS-LS] and [HS-HS]. The relative stability of the [LS-HS] state increases with increasing ligand field strength of the bis(catecholate) bridging ligand in the order 1 < 2 < 3. The bromo substituents of Br4spiro4- increase the ligand field strength relative to spiro4-, while the stronger ligand field provided by thea4- arises from extension of the overlapping π-orbital system across the two catecholate units. This study highlights how SCO behavior in dinuclear complexes can be modulated by the bridging ligand, providing useful insights for the design of molecules that can be interconverted between more than two states.

Docosahexaenoic Acid Is Naturally Concentrated at the sn-2 Position in Triacylglycerols of the Australian Thraustochytrid Aurantiochytrium sp. Strain TC 20.

The Labyrinthulomycetes or Labyrinthulea are a class of protists that produce a network of filaments that enable the cells to glide along and absorb nutrients. One of the main two Labyrinthulea groups is the thraustochytrids, which are becoming an increasingly recognised and commercially used alternate source of long-chain (LC, ≥C20) omega-3 containing oils. This study demonstrates, to our knowledge for the first time, the regiospecificity of the triacylglycerol (TAG) fraction derived from Australian thraustochytrid Aurantiochytrium sp. strain TC 20 obtained using 13C nuclear magnetic resonance spectroscopy (13C NMR) analysis. The DHA present in the TC 20 TAG fraction was determined to be concentrated in the sn-2 position, with TAG (16:0/22:6/16:0) identified as the main species present. The sn-2 preference is similar to that found in salmon and tuna oil, and differs to seal oil containing largely sn-1,3 LC-PUFA. A higher concentration of sn-2 DHA occurred in the thraustochytrid TC 20 oil compared to that of tuna oil.

DFT Prediction and Experimental Investigation of Valence Tautomerism in Cobalt-Dioxolene Complexes.

The family of complexes of general formula [Co(Me ntpa)(Xdiox)]+ (tpa = tris(2-pyridylmethyl)amine, n = 0-3 corresponds to successive methylation of the 6-position of the pyridine rings; X = Br4, Cl4, H4, 3,5-Me2, 3,5- tBu2; diox = dioxolene) was investigated by density functional theory (DFT) calculations to predict the likelihood of valence tautomerism (VT). The OPBE functional with relativistic and solvent corrections allowed accurate reproduction of trends in spin-state energetics, affording the prediction of VT in complex [Co(Me3tpa)(Br4diox)]+ (1+). One-electron oxidation of neutral precursor [CoII(Me3tpa)(Br4cat)] (1) enabled isolation of target compounds 1(PF6) and 1(BPh4). Solution variable-temperature UV-vis absorption and Evans method magnetic susceptibility data confirm DFT predictions that 1+ exists in a temperature-dependent valence tautomeric equilibrium between low-spin Co(III)-catecholate and high-spin Co(II)-semiquinonate forms. The solution VT transition temperature of 1+ is solvent-tunable with critical temperatures in the range of 291-359 K for the solvents measured. Solid-state magnetic susceptibility measurements of 1(PF6) and 1(BPh4) reveal the onset of VT transitions above room temperature.

Visible-Light-Driven "On"/"Off" Photochromism of a Polyoxometalate Diarylethene Coordination Complex.

Herein we report the first photochromic polyoxometalate (POM)-based diarylethene (DAE) coordination complex, prepared by ligation of two cobalt(III)-incorporated borotungstates [BIIIWVI11O39CoIII]6- with the ditopic pyridyl-containing diarylethene (C25H16N2F6S2). The solution-state composition, structure, and stability of the assembly were probed using 1H and 19F nuclear magnetic resonance spectroscopy (NMR), electrospray ionization quadrupolar time-of-flight mass spectrometry (ESI-QTOF-MS), ultraviolet-visible spectroscopy (UV-vis), and small-angle X-ray scattering (SAXS), revealing that the complex self-organizes to adopt a molecular dumbbell structure due to electrostatic and steric considerations. This conformation is a prerequisite for the photocyclization reaction. The assembly was found to be switchable between two states using visible light due to the perturbation of the DAE electronic structure on coordination to the POM. We present photophysical data, including the reaction quantum efficiency of the molecular switch in both directions measured using a custom-built quantum yield determination setup in addition to fatigue resistance on prolonged irradiation.

Synthesis and X-Ray Crystallographic Characterisation of Frustum-Shaped Ligated [Cu18 H16 (DPPE)6 ]2+ and [Cu16 H14 (DPPA)6 ]2+ Nanoclusters and Studies on Their H2 Evolution Reactions.

We report new structural motifs for Cu nanoclusters that conceptually represent seed crystals for large face-centred cubic (FCC) crystal growth. Kinetically controlled syntheses, high resolution mass spectrometry experiments for determination of the dication formulae and crystallographic characterisation were carried out for [Cu18 H16 (DPPE)6 ][BF4 ][Cl] (DPPE=bis(diphenylphosphino)ethane) and [Cu16 H14 (DPPA)6 ][(BF4 )2 ] (DPPA=bis(diphenylphosphino)amine) polyhydrido nanoclusters, which feature the unprecedented bifrustum and frustum metal-core architecture in metal nanoclusters. The Cu18 nanocluster contains two Cu9 frustum cupolae and the Cu16 nanocluster has one Cu9 frustum cupola and a Cu7 distorted hexagonal-shape base. Gas-phase experiments revealed that both Cu18 H16 and Cu16 H14 cores can spontaneously release H2 upon removal of one bisphosphine capping ligand.

Synthesis, Structural Characterization, and Gas-Phase Unimolecular Reactivity of Bis(diphenylphosphino)amino Copper Hydride Nanoclusters [Cu3(X)(µ3-H)((PPh2)2NH)3](BF4), Where X = µ2-Cl and µ3-BH4.

An electrospray ionization mass spectrometry (ESI-MS) survey of the types of cationic copper clusters formed from an acetonitrile solution containing a 1:1:20 mixture of tetrakis(acetonitrile)copper(I) tetrafluoroborate [Cu(MeCN)4(BF4)], bis(diphenylphosphino)amine (dppa = (Ph2P)2NH = L), and NaBH4 revealed a major peak, which based on both the accurate masses and isotope distribution was assigned as [Cu3(BH4)(H)(L)3]+. This prompted synthetic efforts resulting in isolation of the dppa ligated trinuclear copper hydride nanoclusters, [Cu3(µ2-Cl)(µ3-H)(L)3](BF4) and [Cu3(µ3-BH4)(µ3-H)(L)3](BF4), which were subsequently structurally characterized using high resolution ESI-MS, X-ray crystallography, NMR, and IR spectroscopy. The X-ray structures reveal a common structural feature of the cation, in which the three copper(I) ions adopt a planar trinuclear Cu3 geometry coordinated on the bottom face by a µ3-hydride and surrounded by three dppa ligands. ESI-MS of [Cu3(µ2-Cl)(µ3-H)(L)3](BF4) and [Cu3(µ3-BH4)(µ3-H)(L)3](BF4) produces [Cu3(µ2-Cl)(µ3-H)(L)3]+ and [Cu3(µ3-BH4)(µ3-H)(L)3]+. The unimolecular gas-phase ion chemistry of these cations was examined under multistage tandem mass spectrometry conditions using collision-induced dissociation (CID). CID of both cations proceeds via ligand loss to give [Cu3(µ3-H)(X)(L)2]+, which is in competition with BH3 loss in the case of X = BH4. DFT calculations on the fragmentation of [Cu3(µ3-BH4)(µ3-H)(LMe)3]+ suggest that BH3 loss produces the hitherto elusive [Cu3(µ3-H)(µ2-H)(LMe)3]+, which undergoes further fragmentation via H2 loss. CID of the deuterium labeled cluster [Cu3(µ3-D)(µ3-BD4)(L)3]+ reveals that the competing losses of ligand and BD3 yield [Cu3(µ3-BD4)(µ3-D)(L)2]+ and [Cu3(D)2(L)3]+ as primary products, which subsequently fragment via further losses of BD3 or a ligand to give [Cu3(D)2(L)2]+. The coordinated hydrides in the latter ion are activated toward elimination of D2 to give [Cu3(L)2]+. Loss of HD and 2HD are minor channels, consistent with higher DFT predicted endothermicities to form [Cu3(D)(L)(L-H)]+ and [Cu3(L-H)2]+.

Synthesis, structural characterization, and gas-phase unimolecular reactivity of the silver hydride nanocluster [Ag3((PPh2)2CH2)3(µ3-H)](BF4)2.

A bis(diphenylphosphino)methane-ligated trinuclear silver hydride nanocluster, [Ag3((Ph2P)2CH2)3(µ3-H)](BF4)2, featuring three silver(I) ions coordinated to a µ3-hydride, and its deuteride analogue, [Ag3((Ph2P)2CH2)3(µ3-D)](BF4)2, have been isolated and structurally characterized using electrospray ionization mass spectrometry (ESI-MS), X-ray crystallography, NMR and IR spectroscopy. The position of the deuteride in [Ag3((Ph2P)2CH2)3(µ3-D)](BF4)2 was determined by neutron diffraction. ESI-MS of [Ag3L3(µ3-H/D)](BF4)2 [L = ((Ph2P)2CH2)2] produces [Ag3L3(µ3-H/D)](2+) and [Ag3L3(µ3-H/D)(BF4)](+). A rich gas-phase ion chemistry of [Ag3L3(µ3-H/D)](2+) is observed under conditions of collision-induced dissociation (CID) and electron-capture dissociation (ECD). CID gives rise to the following complementary ion pairs: [Ag3L2](+) and [L+(H/D)](+); [Ag2(H/D)L2](+) and [AgL](+); [Ag2(H/D)L](+) and [AgL2](+). ECD gives rise to a number of dissociation channels including loss of the bis(phosphine) ligand, fragmentation of a coordinated bis(phosphine) ligand via C-P bond activation, and loss of a hydrogen (deuterium) atom with concomitant formation of [Ag3L3](+). Under CID conditions, [Ag3L3(µ3-H/D)(BF4)](+) fragments via ligand loss, the combined loss of a ligand and [H,B,F4], and cluster fragmentation to give [Ag2(BF4)L2](+) and [Ag2(L-H)L](+) [where (L-H) = (Ph2P)2CH(-)].

Ending aging in super glassy polymer membranes.

Aging in super glassy polymers such as poly(trimethylsilylpropyne) (PTMSP), poly(4-methyl-2-pentyne) (PMP), and polymers with intrinsic microporosity (PIM-1) reduces gas permeabilities and limits their application as gas-separation membranes. While super glassy polymers are initially very porous, and ultra-permeable, they quickly pack into a denser phase becoming less porous and permeable. This age-old problem has been solved by adding an ultraporous additive that maintains the low density, porous, initial stage of super glassy polymers through absorbing a portion of the polymer chains within its pores thereby holding the chains in their open position. This result is the first time that aging in super glassy polymers is inhibited whilst maintaining enhanced CO2 permeability for one year and improving CO2/N2 selectivity. This approach could allow super glassy polymers to be revisited for commercial application in gas separations.

Gladiolin produced by pathogenic Burkholderia synergizes with amphotericin B through membrane lipid rearrangements.

Amphotericin B (AmpB) is an effective but toxic antifungal drug. Thus, improving its activity/toxicity relationship is of interest. AmpB disrupts fungal membranes by two proposed mechanisms: ergosterol sequestration from the membrane and pore formation. Whether these two mechanisms operate in conjunction and how they could be potentiated remains to be fully understood. Here, we report that gladiolin, a polyketide antibiotic produced by Burkholderia gladioli, is a strong potentiator of AmpB and acts synergistically against Cryptococcus and Candida species, including drug-resistant C. auris. Gladiolin also synergizes with AmpB against drug-resistant fungal biofilms, while exerting no mammalian cytotoxicity. To explain the mechanism of synergy, we show that gladiolin interacts with membranes via a previously unreported binding mode for polyketides. Moreover, gladiolin modulates lipid binding by AmpB and, in combination, causes faster and more pronounced lipid rearrangements relative to AmpB alone which include membrane thinning consistent with ergosterol extraction, areas of thickening, pore formation, and increased membrane destruction. These biophysical data provide evidence of a functional interaction between gladiolin and AmpB at the membrane interface. The data further indicate that the two proposed AmpB mechanisms (ergosterol sequestration and pore formation) act in conjunction to disrupt membranes, and that gladiolin synergizes by enhancing both mechanisms. Collectively, our findings shed light on AmpB's mechanism of action and characterize gladiolin as an AmpB potentiator, showing an antifungal mechanism distinct from its proposed antibiotic activity. We shed light on the synergistic mechanism at the membrane, and provide insights into potentiation strategies to improve AmpB's activity/toxicity relationship. IMPORTANCE: Amphotericin B (AmpB) is one of the oldest antifungal drugs in clinical use. It is an effective therapeutic, but it comes with toxicity issues due to the similarities between its fungal target (the membrane lipid ergosterol) and its mammalian counterpart (cholesterol). One strategy to improve its activity/toxicity relationship is by combinatorial therapy with potentiators, which would enable a lower therapeutic dose of AmpB. Here, we report on the discovery of the antibiotic gladiolin as a potentiator of AmpB against several priority human fungal pathogens and fungal biofilms, with no increased toxicity against mammalian cells. We show that gladiolin potentiates AmpB by increasing and accelerating membrane damage. Our findings also provide insights into the on-going debate about the mechanism of action of AmpB by indicating that both proposed mechanisms, extraction of ergosterol from membranes and pore formation, are potentiated by gladiolin.

Constraints within major histocompatibility complex class I restricted peptides: presentation and consequences for T-cell recognition.

Residues within processed protein fragments bound to major histocompatibility complex class I (MHC-I) glycoproteins have been considered to function as a series of "independent pegs" that either anchor the peptide (p) to the MHC-I and/or interact with the spectrum of alphabeta-T-cell receptors (TCRs) specific for the pMHC-I epitope in question. Mining of the extensive pMHC-I structural database established that many self- and viral peptides show extensive and direct interresidue interactions, an unexpected finding that has led us to the idea of "constrained" peptides. Mutational analysis of two constrained peptides (the HLA B44 restricted self-peptide (B44DPalpha-EEFGRAFSF) and an H2-D(b) restricted influenza peptide (D(b)PA, SSLENFRAYV) demonstrated that the conformation of the prominently exposed arginine in both peptides was governed by interactions with MHC-I-orientated flanking residues from the peptide itself. Using reverse genetics in a murine influenza model, we revealed that mutation of an MHC-I-orientated residue (SSLENFRAYV --> SSLENARAYV) within the constrained PA peptide resulted in a diminished cytotoxic T lymphocyte (CTL) response and the recruitment of a limited pMHC-I specific TCR repertoire. Interactions between individual peptide positions can thus impose fine control on the conformation of pMHC-I epitopes, whereas the perturbation of such constraints can lead to a previously unappreciated mechanism of viral escape.

Highly aqueously stable C60-polymer nanoparticles with excellent photodynamic property for potential cancer treatment.

Fullerenes are a class of carbon nanomaterials that find a wide range of applications in biomedical fields, especially for photodynamic cancer therapy because of its photosensitive effect. However, hydrophobic fullerenes can only be dispersed in organic solvents which hinders their biomedical applications. Here, we report a facile method to prepare highly water-dispersible fullerene (C60)-polymer nanoparticles with hydrodynamic sizes of 50-70 nm. Hydrophilic random copolymers containing different ratios of polyethylene glycol methyl ether methacrylate and 2-aminoethylmethacrylamide were synthesized for conjugating with C60 molecules through efficient nucleophilic Michael addition reaction between amine groups from hydrophilic polymer and carbon-carbon double bonds from C60. As a result, the amphiphilic C60-polymer conjugates could be well dispersed and nano-assembled in water with a C60 concentration as high as 7.8 mg/mL, demonstrating a significant improvement for the solubility of C60 in an aqueous system. Owing to the high C60 content, the C60-polymer nanoparticles showed a strong photodynamic therapy effect on human lung cancer cells (A549) under light irradiation (450 nm) in both 2D cell culture and 3D spheroid culture, while demonstrating ignorable cytotoxicity under dark. This highly efficient and convenient method to prepare water-dispersible C60-polymer conjugates may have a great impact on the future biomedical applications of fullerenes.

Conformational stability studies of a stapled hexa-ß3-peptide library.

A library of 14-helical hexa ß(3)-peptides was synthesized in order to determine the influence of sequence variation as well as staple size and location on conformational stability. From this study we show that appropriately stapled hexa-ß(3)-peptides can allow for a number of variations without significant perturbation of the 14-helix.

Protic ionic liquids with fluorous anions: physicochemical properties and self-assembly nanostructure.

A series of 11 new protic ionic liquids with fluorous anions (FPILs) have been identified and their self-assembled nanostructure, thermal phase transitions and physicochemical properties were investigated. To the best of our knowledge this is the first time that fluorocarbon domains have been reported in PILs. The FPILs were prepared from a range of hydrocarbon alkyl and heterocyclic amine cations in combination with the perfluorinated anions heptafluorobutyrate and pentadecafluorooctanoate. The nanostructure of the FPILs was established by using small- and wide-angle X-ray scattering (SAXS and WAXS). In the liquid state many of the FPILs showed an intermediate range order, or self-assembled nanostructure, resulting from segregation of the polar and nonpolar hydrocarbon and fluorocarbon domains of the ionic liquid. In addition, the physicochemical properties of the FPILs were determined including the melting point (T(m)), glass transition (T(g)), devitrification temperature (T(c)), thermal stability and the density ρ, viscosity η, air/liquid surface tension γ(LV), refractive index n(D), and ionic conductivity κ. The FPILs were mostly solids at room temperature, however two examples 2-pyrrolidinonium heptafluorobutyrate (PyrroBF) and pyrrolidinium heptafluorobutyrate (PyrrBF) were liquids at room temperature and all of the FPILs melted below 80 °C. Four of the FPILs exhibited a glass transition. The two liquids at room temperature, PyrroBF and PyrrBF, had a similar density, surface tension and refractive index but their viscosity and ionic conductivity were very different due to dissimilar self-assembled nanostructure.

A saccharide-based binder for efficient polysulfide regulations in Li-S batteries.

The viability of lithium-sulfur batteries as an energy storage technology depends on unlocking long-term cycle stability. Most instability stems from the release and transport of polysulfides from the cathode, which causes mossy growth on the lithium anode, leading to continuous consumption of electrolyte. Therefore, development of a durable cathode with minimal polysulfide escape is critical. Here, we present a saccharide-based binder system that has a capacity for the regulation of polysulfides due to its reducing properties. Furthermore, the binder promotes the formation of viscoelastic filaments during casting which endows the sulfur cathode with a desirable web-like microstructure. Taken together this leads to 97% sulfur utilisation with a cycle life of 1000 cycles (9 months) and capacity retention (around 700 mAh g-1 after 1000 cycles). A pouch cell prototype with a specific energy of up to 206 Wh kg-1 is produced, demonstrating the promising potential for practical applications.

Development of a Brassica napus (Canola) Crop Containing Fish Oil-Like Levels of DHA in the Seed Oil.

Plant seeds have long been promoted as a production platform for novel fatty acids such as the ω3 long-chain (≥ C20) polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) commonly found in fish oil. In this article we describe the creation of a canola (Brassica napus) variety producing fish oil-like levels of DHA in the seed. This was achieved by the introduction of a microalgal/yeast transgenic pathway of seven consecutive enzymatic steps which converted the native substrate oleic acid to α-linolenic acid and, subsequently, to EPA, docosapentaenoic acid (DPA) and DHA. This paper describes construct design and evaluation, plant transformation, event selection, field testing in a wide range of environments, and oil profile stability of the transgenic seed. The stable, high-performing event NS-B50027-4 produced fish oil-like levels of DHA (9-11%) in open field trials of T3 to T7 generation plants in several locations in Australia and Canada. This study also describes the highest seed DHA levels reported thus far and is one of the first examples of a deregulated genetically modified crop with clear health benefits to the consumer.

A community-built calibration system: The case study of quantification of metabolites in grape juice by qNMR spectroscopy.

Nuclear Magnetic Resonance (NMR) is an analytical technique extensively used in almost every chemical laboratory for structural identification. This technique provides statistically equivalent signals in spite of using spectrometer with different hardware features and is successfully used for the traceability and quantification of analytes in food samples. Nevertheless, to date only a few internationally agreed guidelines have been reported on the use of NMR for quantitative analysis. The main goal of the present study is to provide a methodological pipeline to assess the reproducibility of NMR data produced for a given matrix by spectrometers from different manufacturers, with different magnetic field strengths, age and hardware configurations. The results have been analyzed through a sequence of chemometric tests to generate a community-built calibration system which was used to verify the performance of the spectrometers and the reproducibility of the predicted sample concentrations.

Synthesis and structure-activity relationships of bengazole A analogs.

Analogs of the potent antifungal agent, bengazole A, were prepared and evaluated against Candida spp. in both microbroth dilution and disk diffusion assays.

Solvation Effects on the Permeation and Aging Performance of PIM-1-Based MMMs for Gas Separation.

Membranes are particularly attractive for lowering the energy intensity of separations as they eliminate phase changes. While many tantalizing polymers are known, limitations in selectivity and stability slightly preclude further development. Mixed-matrix membranes may address these shortcomings. Key to their realization is the intimate mixing between the polymer and the additive to eliminate nonselective transport, improve selectivity, and resist physical aging. Polymers of intrinsic microporosity (PIMs) have inherently promising gas transport properties. Here, we show that porous additives can improve transport and resist aging in PIM-1. We develop a simple, low-cost, and scalable hyper-cross-linked polymer (poly-dichloroxylene, pDCX), which was hydroxylated to form an intimate mixture with the polar PIM-1. Solvent variation allowed control of physical aging rates and improved selectivity for smaller gases. This detailed study has allowed many interactions within mixed matrix membranes to be directly elucidated and presents a practical means to stabilize porous polymers for separation applications.

Structural characterization and gas-phase studies of the [Ag10H8(L)6]2+ nanocluster dication.

The reactions between silver salts and borohydrides produce a rich set of products that range from discrete mononuclear compounds through to silver nanoparticles and colloids. Previous studies using electrospray ionization mass spectrometry (ESI-MS) to track the cationic products in solutions containing sodium borohydride, silver(i) tetrafluoroborate and the bisphosphine ligands, L, bis(diphenylphosphino)methane (dppm) and bis(diphenylphosphino)amine (dppa) have identified the dications [Ag10H8(L)6]2+. Here we isolate and structurally characterize [Ag10H8(dppa)6](BF4)2, and [Ag10H8(dppa)6](NO3)2via X-ray crystallography. Both dications have nearly identical structural features consisting of a Ag10 scaffold with the atoms lying on vertices of a bicapped square antiprism. DFT calculations were carried out to suggest potential sites for the hydrides. Ion-mobility mass spectrometry experiments revealed that [Ag10H8(dppa)6]2+ and [Ag10H8(dppm)6]2+ have similar collision cross sections, while multistage mass spectrometry experiments were used to compare their unimolecular gas-phase chemistry. Although the same initial sequential ligand loss followed by cluster fission and H2 evolution is observed, the more acidic N-H of the dppa provides a more labile H for H2 loss and H/D scrambling processes as revealed by isotope labelled experiments.

A synthetic resilin is largely unstructured.

Proresilin is the precursor protein for resilin, an extremely elastic, hydrated, cross-linked insoluble protein found in insects. We investigated the secondary-structure distribution in solution of a synthetic proresilin (AN16), based on 16 units of the consensus proresilin repeat from Anopheles gambiae. Raman spectroscopy was used to verify that the secondary-structure distributions in cross-linked AN16 resilin and in AN16 proresilin are similar, and hence that solution techniques (such as NMR and circular dichroism) may be used to gain information about the structure of the cross-linked solid. The synthetic proresilin AN16 is an intrinsically unstructured protein, displaying under native conditions many of the characteristics normally observed in denatured proteins. There are no apparent alpha-helical or beta-sheet features in the NMR spectra, and the majority of backbone protons and carbons exhibit chemical shifts characteristic of random-coil configurations. Relatively few peaks are observed in the nuclear Overhauser effect spectra, indicating that overall the protein is dynamic and unstructured. The radius of gyration of AN16 corresponds to the value expected for a denatured protein of similar chain length. This high degree of disorder is also consistent with observed circular dichroism and Raman spectra. The temperature dependences of the NH proton chemical shifts were also measured. Most values were indicative of protons exposed to water, although smaller dependences were observed for glycine and alanine within the Tyr-Gly-Ala-Pro sequence conserved in all resilins found to date, which is the site of dityrosine cross-link formation. This result implies that these residues are involved in hydrogen bonds, possibly to enable efficient self-association and subsequent cross-linking. The beta-spiral model for elastic proteins, where the protein is itself shaped like a spring, is not supported by the results for AN16. Both the random-network elastomer model and the sliding beta-turn model are consistent with the data. The results indicate a flat energy landscape for AN16, with very little energy required to switch between conformations. This ease of switching is likely to lead to the extremely low energy loss on deformation of resilin.