Evidence for Acyl Homoserine Lactone Signal Production in Bacteria Associated with Marine Sponges

We report for the first time the production of acyl homoserine lactones (AHLs) by bacteria associated with marine sponges. Given the involvement of AHLs in bacterial colonization of many higher organisms, we speculate that such quorum sensing signals could play a part in interactions between sponges and the dense bacterial communities living within them.     

The effect of temperature on larval pre-settlement duration and metamorphosis for the sponge, <Emphasis Type="Italic">Rhopaloeides odorabile</Emphasis>

Rising sea temperatures may potentially affect the dispersive larval phase of sessile marine invertebrates with consequences for the viability of adult populations. This study demonstrated that the planktonic larvae of Rhopaloeides odorabile, a common Great Barrier Reef sponge, survived and metamorphosed when exposed to temperatures up to 9°C above the annual maximum (~29°C). Planktonic larval duration of 54 h, at ambient temperatures (~28°C), were reduced to 18 h for larvae exposed to elevated temperatures (32–36°C). Moreover, at ambient temperatures larvae began metamorphosing after 12 h, but at 32–36°C this reduced to only 2 h. Larvae survived and could still metamorphose at temperatures as high as 38°C, but were no longer functional at 40°C. These results imply that predicted increases in sea surface temperature may reduce planktonic larval duration and dispersal capabilities, thereby contributing to population subdivision of the species.     

Characterisation of human peroxisomal 2,4-dienoyl-CoA reductase12

Based on the primary structure of the rat peroxisomal 2,4-dienoyl-CoA reductase (M. Fransen, P.P. Van Veldhoven, S. Subramani, Biochem. J. 340 (1999) 561–568), the cDNA of the human counterpart was cloned. It contained an open reading frame of 878 bases encoding a protein of 291 amino acids (calculated molecular mass 30?778 Da), being 83% identical to the rat reductase. The gene, encompassing nine exons, is located at chromosome 16p13. Bacterially expressed poly(His)-tagged reductase was active not only towards short and medium chain 2,4-dienoyl-CoAs, but also towards 2,4,7,10,13,16,19-docosaheptaenoyl-CoA. Hence, the reductase does not seem to constitute a rate limiting step in the peroxisomal degradation of docosahexaenoic acid. The reduction of docosaheptaenoyl-CoA, however, was severely decreased in the presence of albumin.     

Ovocalyxin-32, a novel chicken eggshell matrix protein. isolation, amino acid sequencing, cloning, and immunocytochemical localization

The eggshell is a highly ordered structure resulting from the deposition of calcium carbonate concomitantly with an organic matrix upon the eggshell membranes. Mineralization takes place in an acellular uterine fluid, which contains the ionic and matrix precursors of the eggshell. We have identified a novel 32-kDa protein, ovocalyxin-32, which is expressed at high levels in the uterine and isthmus regions of the oviduct, and concentrated in the eggshell. Sequencing of peptides derived from the purified protein allowed expressed sequence tag sequences to be identified that were assembled to yield a full-length composite sequence whose conceptual translation product contained the complete amino acid sequence of ovocalyxin-32. Data base searches revealed that ovocalyxin-32 has limited identity (32%) to two unrelated proteins: latexin, a carboxypeptidase inhibitor expressed in the rat cerebral cortex and mast cells, and a skin protein, which is encoded by a retinoic acid receptor-responsive gene, TIG1. High level expression of ovocalyxin-32 was limited to the isthmus and uterus tissue, where immunocytochemistry at the light and electron microscope levels demonstrated that ovocalyxin-32 is secreted by surface epithelial cells. In the eggshell, ovocalyxin-32 localizes to the outer palisade layer, the vertical crystal layer, and the cuticle of the eggshell, in agreement with its demonstration by Western blotting at high levels in the uterine fluid during the termination phase of eggshell formation. Ovocalyxin-32 is therefore identified as a novel protein synthesized in the distal oviduct where hen eggshell formation occurs.     

Fouling deterrence on the bivalve shell Mytilus galloprovincialis: a physical phenomenon?

The physical nature of fouling deterrence by the mussel Mytilus galloprovincialis was investigated using high-resolution biomimics of the bivalve surface. The homogeneous microtextured surface of M. galloprovincialis (1.94 +/- 0.03 microm), the smooth surface of the bivalve Amusium balloti (0 microm), and moulds of these surfaces (biomimics) were compared with controls of smooth (0 microm) and sanded moulds, (55.4 +/- 2.7 microm) and PVC strips (0 microm) in a 12-week field trial. The shell and mould of M. galloprovincialis were fouled by significantly fewer species and had significantly less total fouling cover than the shell and mould of A. balloti over a 12-week period. However, the major effects were between surfaces with and without microtopography. Surface microtopography, be it structured as in the case of M. galloprovincialis shell and mould, or random as in the case of the sanded mould, had a lower cover of fouling organisms than treatments without microtopography after 6 weeks. There was also no difference between the effect of the M. galloprovincialis mould and the sanded mould. The strong fouling deterrent effects of both these surfaces diminished rapidly after 6 to 8 weeks while that of M. galloprovincialis shell remained intact for the duration of the experiment suggesting factors in addition to surface microtopography contribute to fouling deterrence.     

Stoichiometry of Heteromeric BAFF and APRIL Cytokines Dictates Their Receptor Binding and Signaling Properties

The closely related TNF family ligands B cell activation factor (BAFF) and a proliferation-inducing ligand (APRIL) serve in the generation and maintenance of mature B-lymphocytes. Both BAFF and APRIL assemble as homotrimers that bind and activate several receptors that they partially share. However, heteromers of BAFF and APRIL that occur in patients with autoimmune diseases are incompletely characterized. The N and C termini of adjacent BAFF or APRIL monomers are spatially close and can be linked to create single-chain homo- or hetero-ligands of defined stoichiometry. Similar to APRIL, heteromers consisting of one BAFF and two APRILs (BAA) bind to the receptors B cell maturation antigen (BCMA), transmembrane activator and CAML interactor (TACI) but not to the BAFF receptor (BAFFR). Heteromers consisting of one APRIL and two BAFF (ABB) bind to TACI and BCMA and weakly to BAFFR in accordance with the analysis of the receptor interaction sites in the crystallographic structure of ABB. Receptor binding correlated with activity in reporter cell line assays specific for BAFFR, TACI, or BCMA. Single-chain BAFF (BBB) and to a lesser extent single-chain ABB, but not APRIL or single-chain BAA, rescued BAFFR-dependent B cell maturation in BAFF-deficient mice. In conclusion, BAFF-APRIL heteromers of different stoichiometries have distinct receptor-binding properties and activities. Based on the observation that heteromers are less active than BAFF, we speculate that their physiological role might be to down-regulate BAFF activity.     

Comparing the potential production and value of high-energy liquid fuels and protein from marine and freshwater macroalgae

The biomass production and biochemical properties of marine and freshwater species of green macroalgae (multicellular algae), cultivated in outdoor conditions, were evaluated to assess the potential conversion into high-energy liquid biofuels, specifically biocrude and biodiesel and the value of these products. Biomass productivities were typically two times higher for marine macroalgae (8.5–11.9 g m⁻² d⁻¹, dry weight) than for freshwater macroalgae (3.4–5.1 g m⁻² d⁻¹, dry weight). The biochemical compositions of the species were also distinct, with higher ash content (25.5–36.6%) in marine macroalgae and higher calorific value (15.8–16.4 MJ kg⁻¹) in freshwater macroalgae. Lipid content was highest for freshwater Oedogonium and marine Derbesia. Lipids are a critical organic component for biocrude production by hydrothermal liquefaction (HTL) and the theoretical biocrude yield was therefore highest for Oedogonium (17.7%, dry weight) and Derbesia (16.2%, dry weight). Theoretical biocrude yields were also higher than biodiesel yields for all species due to the conversion of the whole organic component of biomass, including the predominant carbohydrate fraction. However, all marine species had higher biomass productivities and therefore had higher projected biocrude productivities than freshwater species, up to 7.1 t of biocrude ha⁻¹ yr⁻¹ for Derbesia. The projected value of the six macroalgae was increased by 45–77% (up to US$7700 ha⁻¹ yr⁻¹) through the extraction of protein prior to the conversion of the residual biomass to biocrude. This study highlights the importance of optimizing biomass productivities for high-energy fuels and targeting additional coproducts to increase value.     

Structure of the SthK Carboxy-Terminal Region Reveals a Gating Mechanism for Cyclic Nucleotide-Modulated Ion Channels

Cyclic nucleotide-sensitive ion channels are molecular pores that open in response to cAMP or cGMP, which are universal second messengers. Binding of a cyclic nucleotide to the carboxyterminal cyclic nucleotide binding domain (CNBD) of these channels is thought to cause a conformational change that promotes channel opening. The C-linker domain, which connects the channel pore to this CNBD, plays an important role in coupling ligand binding to channel opening. Current structural insight into this mechanism mainly derives from X-ray crystal structures of the C-linker/CNBD from hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels. However, these structures reveal little to no conformational changes upon comparison of the ligand-bound and unbound form. In this study, we take advantage of a recently identified prokaryote ion channel, SthK, which has functional properties that strongly resemble cyclic nucleotide-gated (CNG) channels and is activated by cAMP, but not by cGMP. We determined X-ray crystal structures of the C-linker/CNBD of SthK in the presence of cAMP or cGMP. We observe that the structure in complex with cGMP, which is an antagonist, is similar to previously determined HCN channel structures. In contrast, the structure in complex with cAMP, which is an agonist, is in a more open conformation. We observe that the CNBD makes an outward swinging movement, which is accompanied by an opening of the C-linker. This conformation mirrors the open gate structures of the K_v1.2 channel or MthK channel, which suggests that the cAMP-bound C-linker/CNBD from SthK represents an activated conformation. These results provide a structural framework for better understanding cyclic nucleotide modulation of ion channels, including HCN and CNG channels.