The significance of organic carbon and nutrient export from peatland-dominated landscapes subject to disturbance, a stoichiometric perspective

The terrestrial-aquatic interface is a crucial environment in which to consider the fate of exported terrestrial carbon in the aquatic system. Here the fate of dissolved organic carbon (DOC) may be controlled by nutrient availability. However, peat-dominated headwater catchments are normally of low nutrient status and thus there is little data on how DOC and nutrient export co-varies. We present nutrient and DOC data for two UK catchments dominated by peat headwaters. One, Whitelee, is undergoing development for Europe's largest windfarm. Glen Dye by comparison is relatively undisturbed. At both sites there are significant linear relationships between DOC and soluble reactive phosphorus and nitrate concentrations in the drainage waters. However, inter-catchment differences exist. Changes in the pattern of nutrient and carbon export at Whitelee reveal that landscape disturbance associated with windfarm development impacts the receiving waters, and that nutrient export does not increase in a stoichiometric manner that will promote increase in microbial biomass but rather supports aquatic respiration. In turn greater CO2 efflux may prevail. Hence disturbance of terrestrial carbon stores may impact the both the aquatic and gaseous carbon cycle. We suggest estimates of aquatic carbon export should inform the decision-making process prior to development in ecosystems and catchments with high terrestrial carbon storage

The significance of organic carbon and nutrient export from peatland-dominated landscapes subject to disturbance

International audienceThe terrestrial-aquatic interface is a crucial environment in which to consider the fate of exported terrestrial carbon in the aquatic system. To a large extent the fate of dissolved organic carbon (DOC) may be controlled by nutrient availability. However, peat-dominated headwater catchments are normally considered of low nutrient status and thus there is little data on the interaction of DOC and nutrients. Here we present nutrient and DOC data exported from two UK catchments, both dominated by peat headwaters, but of differing land-use. Glen Dye is a moorland with no trees; Whitelee has partially forested peats and peaty podzols, and is now undergoing development to host Europe's largest on-shore windfarm, the Whitelee Windfarm. There are significant linear relationships between DOC and soluble reactive phosphorus and nitrate concentrations in the drainage waters, but inter-catchment differences exist. Changes in the pattern of nutrient and carbon export in Whitelee suggest that disturbance of peatlands soils can impact the receiving water and that nutrient export does not increase in a stoichiometric manner that will promote increase in biomass. As such the changes are more likely to cause increased aquatic respiration, and thus lead to higher dissolved CO2 concentrations (and therefore CO2 efflux). Hence disturbance of terrestrial carbon stores may also impact the gaseous carbon cycle. Confirming the source of carbon and nutrients in these study sites is not possible. However, nearby 14C measurements are in keeping with other published literature values from similar sites which show C in DOM exported from peatlands is predominantly modern, and thus supports an interpretation that nutrients, additional to carbon, are derived from shallow soils. Estimates of organic carbon loss from Whitelee catchments to the drainage waters suggest a system where losses are approaching likely sequestration rates. We suggest such sequestration assessment should inform the decision-making tools required prior to development of terrestrial carbon stores

The serine protease domain of MASP-3: enzymatic properties and crystal structure in complex with ecotin.

International audienceMannan-binding lectin (MBL), ficolins and collectin-11 are known to associate with three homologous modular proteases, the MBL-Associated Serine Proteases (MASPs). The crystal structures of the catalytic domains of MASP-1 and MASP-2 have been solved, but the structure of the corresponding domain of MASP-3 remains unknown. A link between mutations in the MASP1/3 gene and the rare autosomal recessive 3MC (Mingarelli, Malpuech, Michels and Carnevale,) syndrome, characterized by various developmental disorders, was discovered recently, revealing an unexpected important role of MASP-3 in early developmental processes. To gain a first insight into the enzymatic and structural properties of MASP-3, a recombinant form of its serine protease (SP) domain was produced and characterized. The amidolytic activity of this domain on fluorescent peptidyl-aminomethylcoumarin substrates was shown to be considerably lower than that of other members of the C1r/C1s/MASP family. The E. coli protease inhibitor ecotin bound to the SP domains of MASP-3 and MASP-2, whereas no significant interaction was detected with MASP-1, C1r and C1s. A tetrameric complex comprising an ecotin dimer and two MASP-3 SP domains was isolated and its crystal structure was solved and refined to 3.2 Å. Analysis of the ecotin/MASP-3 interfaces allows a better understanding of the differential reactivity of the C1r/C1s/MASP protease family members towards ecotin, and comparison of the MASP-3 SP domain structure with those of other trypsin-like proteases yields novel hypotheses accounting for its zymogen-like properties in vitro

Complement Factor H-Related Proteins CFHR2 and CFHR5 Represent Novel Ligands for the Infection-Associated CRASP Proteins of Borrelia burgdorferi

Background: One virulence property of Borrelia burgdorferi is its resistance to innate immunity, in particular to complement-mediated killing. Serum-resistant B. burgdorferi express up to five distinct complement regulator-acquiring surface proteins (CRASP) which interact with complement regulator factor H (CFH) and factor H-like protein 1 (FHL1) or factor H-related protein 1 (CFHR1). In the present study we elucidate the role of the infection-associated CRASP-3 and CRASP-5 protein to serve as ligands for additional complement regulatory proteins as well as for complement resistance of B. burgdorferi. Methodology/Principal Findings: To elucidate whether CRASP-5 and CRASP-3 interact with various human proteins, both borrelial proteins were immobilized on magnetic beads. Following incubation with human serum, bound proteins were eluted and separated by Glycine-SDS-PAGE. In addition to CFH and CFHR1, complement regulators CFHR2 and CFHR5 were identified as novel ligands for both borrelial proteins by employing MALDI-TOF. To further assess the contributions of CRASP-3 and CRASP-5 to complement resistance, a serum-sensitive B. garinii strain G1 which lacks all CFH-binding proteins was used as a valuable model for functional analyses. Both CRASPs expressed on the B. garinii outer surface bound CFH as well as CFHR1 and CFHR2 in ELISA. In contrast, live B. garinii bound CFHR1, CFHR2, and CFHR5 and only miniscute amounts of CFH as demonstrated by serum adsorption assays and FACS analyses. Further functional analysis revealed that upon NHS incubation, CRASP-3 or CRASP-5 expressing borreliae were killed by complement. Conclusions/Significance: In the absence of CFH and the presence of CFHR1, CFHR2 and CFHR5, assembly and integration of the membrane attack complex was not efficiently inhibited indicating that CFH in co-operation with CFHR1, CFHR2 and CFHR5 supports complement evasion of B. burgdorferi

Recombinant human complement component C2 produced in a human cell line restores the classical complement pathway activity in-vitro: an alternative treatment for C2 deficiency diseases

Background: Complement C2 deficiency is the most common genetically determined complete complement deficiency and is associated with a number of diseases. Most prominent are the associations with recurrent serious infections in young children and the development of systemic lupus erythematosus (SLE) in adults. The links with these diseases reflect the important role complement C2 plays in both innate immunity and immune tolerance. Infusions with normal fresh frozen plasma for the treatment of associated disease have demonstrated therapeutic effects but so far protein replacement therapy has not been evaluated. Results: Human complement C2 was cloned and expressed in a mammalian cell line. The purity of recombinant human C2 (rhC2) was greater than 95% and it was characterized for stability and activity. It was sensitive to C1s cleavage and restored classical complement pathway activity in C2-deficient serum both in a complement activation ELISA and a hemolytic assay. Furthermore, rhC2 could increase C3 fragment deposition on the human pathogen Streptococcus pneumoniae in C2-deficient serum to levels equal to those with normal serum. Conclusions: Taken together these data suggest that recombinant human C2 can restore classical complement pathway activity and may serve as a potential therapeutic for recurring bacterial infections or SLE in C2-deficient patients