Changes in zinc ligation promote remodeling of the active site in the zinc hydrolase superfamily

The zinc hydrolase superfamily is a group of divergently related proteins that are predominantly enzymes with a zinc-based catalytic mechanism. The common structural scaffold of the superfamily consists of an eight-stranded β-sheet flanked by six α-helices. Previous analyses, while acknowledging the likely divergent origins of leucine aminopeptidase, carboxypeptidase A and the co-catalytic enzymes of the metallopeptidase H clan based on their structural scaffolds, have failed to find any homology between the active sites in leucine aminopeptidase and the metallopeptidase H clan enzymes. Here we show that these two groups of co-catalytic enzymes have overlapping dizinc centers where one of the two zinc atoms is conserved in each group. Carboxypeptidase A and leucine aminopeptidase, on the other hand, no longer share any homologous zinc-binding sites. At least three catalytic zinc-binding sites have existed in the structural scaffold over the period of history defined by available structures. Comparison of enzyme-inhibitor complexes show that major remodeling of the substrate-binding site has occurred in association with each change in zinc ligation in the binding site. These changes involve re-registration and re-orientation of the substrate. Some residues important to the catalytic mechanism are not conserved amongst members. We discuss how molecules acting in trans may have facilitated the mutation of catalytically important residues in the active site in this group.

Novel B-site ordered double perovskite Ba2Bi0.1Sc0.2Co1.7O6-x for highly efficient oxygen reduction reaction

Double perovskite Ba₂Bi_0.1Sc_0.2Co_1.7O_6-x (BBSC) demonstrates low polarization resistance between 600 and 750 °C due to the high oxygen reduction rate of BBSC as reflected by its large D_V and k values, which are derived from the face centered cubic structure and high cobalt content.

Small home ranges and high site fidelity in red knots (Calidris c. canutus) wintering on the Banc d’Arguin, Mauritania

Using automated and manual radio-telemetry and resightings of individual colour-ringed birds, we assessed the daily use of space of red knots Calidris canutus canutus at a tropical wintering area along the Sahara coast, the Banc d Arguin in Mauritania. Confirming earlier suggestions, we found that birds were very faithful to their roosts and that the daily foraging range was small; in the course of several winter months birds used an area of only 2 16 km2 of intertidal area. We found no differences between their movements in daylight and at night. Additionally, individuals seem to return to exactly the same locations in subsequent winters. This pattern is very different from red knots wintering in the temperate Wadden Sea. Here, they readily change roost sites and easily cover areas of about 800 km2 in the course of weeks but, just as in Mauritania, no differences between day and night are apparent. In northern Patagonia and north-western Australia, red knots have range sizes closer to those on the Banc d Arguin, but here they do show differences in space use between day and night. Ecological explanations for these contrasting patterns require further comparative data based on in-depth studies on the predictability of the food base and the presence of diurnal and nocturnal predators.

Densities of individually marked migrants away from the marking site to estimate population sizes : a test with three wader populations

Capsule Population estimates based on the mark–resighting method can be a useful alternative to population-wide counts.

Aims To investigate whether the mark–resighting method can be used as an alternative to counts to estimate the size of wader populations.

Methods Individual colour-marking and subsequent resightings allowed accurate estimates of annual survival for three populations of waders, on which basis we could estimate the actual number of marked birds alive. Densities of marked birds were determined on sites away (2000–4300 km) from the ringing locations expecting marked birds to be randomly distributed among non-marked conspecifics. Population sizes are estimated by combining these densities with the number of marked birds alive.

Results We found indications that the distribution of marked birds was indeed random in the locations away from the site of marking. The estimated population size of Red Knot Calidris canutus canutus was in accordance with the most recent estimates based on counts. Our estimate of the Calidris c. islandica population was somewhat lower, and that of the Bar-tailed Godwit Limosa lapponica taymyrensis population was considerably lower than the latest estimates based on counts.

Conclusion Population estimates based on the mark–resighting method can be a useful alternative for, or addition to, population-wide counts, as long as the assumption of random distribution of marked birds at the reading sites is taken into account. We conclude that the Afro-Siberian Bar-tailed Godwit population has recently decreased in size or has been substantially overestimated during the counts.

Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination

RNA polymerase II (pol II) transcription termination requires co-transcriptional recognition of a functional polyadenylation signal, but the molecular mechanisms that transduce this signal to pol II remain unclear. We show that Yhh1p/Cft1p, the yeast homologue of the mammalian AAUAAA interacting protein CPSF 160, is an RNA-binding protein and provide evidence that it participates in poly(A) site recognition. Interestingly, RNA binding is mediated by a central domain composed of predicted -propeller-forming repeats, which occurs in proteins of diverse cellular functions. We also found that Yhh1p/Cft1p bound specifically to the phosphorylated C-terminal domain (CTD) of pol II in vitro and in a two-hybrid test in vivo. Furthermore, transcriptional run-on analysis demonstrated that yhh1 mutants were defective in transcription termination, suggesting that Yhh1p/Cft1p functions in the coupling of transcription and 3'-end formation. We propose that direct interactions of Yhh1p/Cft1p with both the RNA transcript and the CTD are required to communicate poly(A) site recognition to elongating pol II to initiate transcription termination.

Phenylalanine-544 plays a key role in substrate and inhibitor binding by providing a hydrophobic packing point at the active site of insulin-regulated aminopeptidase

Inhibitors of insulin-regulated aminopeptidase (IRAP) improve memory and are being developed as a novel treatment for memory loss. In this study, the binding of a class of these inhibitors to human IRAP was investigated using molecular docking and site-directed mutagenesis. Four benzopyran-based IRAP inhibitors with different affinities were docked into a homology model of the catalytic site of IRAP. Two 4-pyridinyl derivatives orient with the benzopyran oxygen interacting with the Zn2+ ion and a direct parallel ring-stack interaction between the benzopyran rings and Phe544. In contrast, the two 4-quinolinyl derivatives orient in a different manner, interacting with the Zn2+ ion via the quinoline nitrogen, and Phe544 contributes an edge-face hydrophobic stacking point with the benzopyran moiety. Mutagenic replacement of Phe544 with alanine, isoleucine, or valine resulted in either complete loss of catalytic activity or altered hydrolysis velocity that was substrate-dependent. Phe544 is also important for inhibitor binding, because these mutations altered the Ki in some cases, and docking of the inhibitors into the corresponding Phe544 mutant models revealed how the interaction might be disturbed. These findings demonstrate a key role of Phe544 in the binding of the benzopyran IRAP inhibitors and for optimal positioning of enzyme substrates during catalysis.

A Ramsar wetland in crisis the Coorong, Lower Lakes and Murray Mouth, Australia

The state of global freshwater ecosystems is increasingly parlous with water resource development degrading high-conservation wetlands. Rehabilitation is challenging because necessary increases in environmental flows have concomitant social impacts, complicated because many rivers flow between jurisdictions or countries. Australia's MurrayDarling Basin is a large river basin with such problems encapsulated in the crisis of its Ramsar-listed terminal wetland, the Coorong, Lower Lakes and Murray Mouth. Prolonged drought and upstream diversion of water dropped water levels in the Lakes below sea level (20092010), exposing hazardous acid sulfate soils. Salinities increased dramatically (e.g. South Lagoon of Coorong>200gL-1, cf. modelled natural 80gL-1), reducing populations of waterbirds, fish, macroinvertebrates and littoral plants. Calcareous masses of estuarine tubeworms (Ficopomatus enigmaticus) killed freshwater turtles (Chelidae) and other fauna. Management primarily focussed on treating symptoms (e.g. acidification), rather than reduced flows, at considerable expense (≥AU$2 billion). We modelled a scenario that increased annual flows during low-flow periods from current levels up to one-third of what the natural flow would have been, potentially delivering substantial environmental benefits and avoiding future crises. Realisation of this outcome depends on increasing environmental flows and implementing sophisticated river management during dry periods, both highly contentious options.