Molten globule-like state of peanut lectin monomer retains its carbohydrate specificity - Implications in protein folding and legume lectin oligomerization

A central question in biological chemistry is the minimal structural requirement of a protein that would determine its specificity and activity, the underlying basis being the importance of the entire structural element of a protein with regards to its activity vis a vis the overall integrity and stability of the protein. Although there are many reports on the characterization of protein folding/ unfolding intermediates, with considerable secondary structural elements but substantial loss of tertiary structure, none of them have been reported to show any activity toward their respective ligands. This may be a result of the conditions under which such intermediates have been isolated or due to the importance of specific structural elements for the activity. In this paper we report such an intermediate in the unfolding of peanut agglutinin that seems to retain, to a considerable degree, its carbohydrate binding specificity and activity. This result has significant implications on the molten globule state during the folding pathway(s) of proteins in general and the quaternary association in legume lectins in particular, where precise subunit topology is required for their biologic activities.

Affinity of a galactose-specific legume lectin from Dolichos lablab to adenine revealed by X-ray cystallography

Crystal structure analysis of a galactose-specific lectin from a leguminous food crop Dolichos lablab (Indian lablab beans) has been carried out to obtain insights into its quaternary association and lectin-carbohydrate interactions. The analysis led to the identification of adenine binding sites at the dimeric interfaces of the heterotetrameric lectin. Structural details of similar adenine binding were reported in only one legume lectin, Dolichos biflorus, before this study. Here, we present the structure of the galactose-binding D. lablab lectin at different pH values in the native form and in complex with galactose and adenine. This first structure report on this lectin also provides a high resolution atomic view of legume lectin-adenine interactions. The tetramer has two canonical and two DB58-like interfaces. The binding of adenine, a non-carbohydrate ligand, is found to occur at four hydrophobic sites at the core of the tetramer at the DB58-like dimeric interfaces and does not interfere with the carbohydrate-binding site. To support the crystallographic observations, the adenine binding was further quantified by carrying out isothermal calorimetric titration. By this method, we not only estimated the affinity of the lectin to adenine but also showed that adenine binds with negative cooperativity in solution.

Of plants and pikas: evidence for a climate-mediated decline in forage and cache quality

Background: Animals that hoard food to mediate seasonal deficits in resource availability might be particularly vulnerable to climate-mediated reductions in the quality and accessibility of food during the caching season. Central-place foragers might be additionally impacted by climatic constraints on their already restricted foraging range. Aims: We sought evidence for these patterns in a study of the American pika (Ochotona princeps), a territorial, central-place forager sensitive to climate. Methods: Pika food caches and available forage were re-sampled using historical methods at two long-term study sites, to quantify changes over two decades. Taxa that changed in availability or use were analysed for primary and secondary metabolites. Results: Both sites trended towards warmer summers, and snowmelt trended earlier at the lower latitude site. Graminoid cover increased at each site, and caching trends appeared to reflect available forage rather than primary metabolites. Pikas at the lower latitude site preferred species higher in secondary metabolites, known to provide higher-nutrient winter forage. However, caching of lower-nutrient graminoids increased in proportion with graminoid availability at that site. Conclusions: If our results represent trends in climate, cache quality and available forage, we predict that pikas at the lower latitude site will soon face nutritional deficiencies.

CLXXX. Vegetable Proteins. I. The Proteins of Dolichos Lab Lab

The field bean (Dolichos lab lab ; Tamil name, Mochai ; Kanarese, Avarai) is a legume which is widely cultivated in South India often as a mixed crop with cereals. The kernel of the seed enters into the diet of may South Indian households, and in the Mysore State the seed are used as a delicacy when they are green for over four months in the year. The haulm, husk and pods are commonly used a fodder. As the kernel which is widely used as an article of food and considered to be very nutritious, contains about 24% of protein hitherto uninvestigated and as the quality of protein plays an important role in nutrition, the present work was undertaken.

Visual ecology of Indian carpenter bees II: adaptations of eyes and ocelli to nocturnal and diurnal lifestyles

Most bees are diurnal, with behaviour that is largely visually mediated, but several groups have made evolutionary shifts to nocturnality, despite having apposition compound eyes unsuited to vision in dim light. We compared the anatomy and optics of the apposition eyes and the ocelli of the nocturnal carpenter bee, Xylocopa tranquebarica, with two sympatric species, the strictly diurnal X. leucothorax and the occasionally crepuscular X. tenuiscapa. The ocelli of the nocturnal X. tranquebarica are unusually large (diameter ca. 1 mm) and poorly focussed. Moreover, their apposition eyes show specific visual adaptations for vision in dim light, including large size, large facets and very wide rhabdoms, which together make these eyes 9 times more sensitive than those of X. tenuiscapa and 27 times more sensitive than those of X. leucothorax. These differences in optical sensitivity are surprisingly small considering that X. tranquebarica can fly on moonless nights when background luminance is as low as 10(-5) cd m(-2), implying that this bee must employ additional visual strategies to forage and find its way back to the nest. These strategies may include photoreceptors with longer integration times and higher contrast gains as well as higher neural summation mechanisms for increasing visual reliability in dim light.

Complexes of lanthanide perchlorates with two new O,N,O ligands derived from antipyraldehyde and acetic and benzoic acid hydrazides

Antipyrine is a well known ligand for lanthanides (I). A forage through the organic literature of pyrazolones reveals that the 4-position of antipyrine is amenable to a wide variety of organic reactions. It should thus be possible to introduce suitable functional groups at this position and design new multidentate ligands for metal ions. It is also found that the coordination chemistry of lanthanides is much less well developed and far fewer ligands have been used for complexation with lanthanide ions compared to that of the d-transition metal ions. Keeping these points in view we have reported earlier, complexes of lanthanides with a bidentate ligand N,N-diethyl-antipyrine-4-carboxamide (2). In this communication we report the synthesis of two new ligands from Schiff base condensation of antipyraldehyde and the hydrazides of acetic and benzoic acids and the complexes formed by these hydrazones with lanthanide perchlorates.

Resolution and sensitivity of the eyes of the Asian honeybees Apis florea, Apis cerana and Apis dorsata

Bees of the genus Apis are important foragers of nectar and pollen resources. Although the European honeybee, Apis mellifera, has been well studied with respect to its sensory abilities, learning behaviour and role as pollinators, much less is known about the other Apis species. We studied the anatomical spatial resolution and absolute sensitivity of the eyes of three sympatric species of Asian honeybees, Apis cerana, Apis florea and Apis dorsata and compared them with the eyes of A. mellifera. Of these four species, the giant honeybee A. dorsata (which forages during moonlit nights) has the lowest spatial resolution and the most sensitive eyes, followed by A. mellifera, A. cerana and the dwarf honeybee, A. florea (which has the smallest acceptance angles and the least sensitive eyes). Moreover, unlike the strictly diurnal A. cerana and A. florea, A. dorsata possess large ocelli, a feature that it shares with all dim-light bees. However, the eyes of the facultatively nocturnal A. dorsata are much less sensitive than those of known obligately nocturnal bees such as Megalopta genalis in Panama and Xylocopa tranquebarica in India. The differences in sensitivity between the eyes of A. dorsata and other strictly diurnal Apis species cannot alone explain why the former is able to fly, orient and forage at half-moon light levels. We assume that additional neuronal adaptations, as has been proposed for A. mellifera, M. genalis and X. tranquebarica, might exist in A. dorsata.

Complexes of lanthanide perchlorates with two new " O,N,O " ligands derived from antipyraldehyde and acetic and benzoic acid hydrazides

Antipyrlne is a well known llgand for lanthanldes (i). A forage through the organic literature of pyrazolones reveals that the 4-position of antipyrlne is amenable to a wide variety of organic reactions. It should thus be possible to introduce suitable functional groups at this position and design new multidentate ligands for metal ions. It is also found that the coordination chemistry of lanthanides is much less well developed and far fewer ligands have been used for complexation with lanthanide ions compared to that of the d-transition metal ions.

Unusual structural stability and ligand induced alterations in oligomerization of a galectin

Abstract L-14, a 14-kDa S-type lectin shows the jelly roll tertiary structural fold akin to legume lectins yet, unlike them, it does not dissociate on thermal unfolding. In the absence of ligand L-14 displays denaturation transitions corresponding to tetrameric and octameric entities. The presence of complementary ligand reduces the association of L-14, which is in stark contrast with legume lectins where no alterations in quaternary structures are brought about by saccharides. From the magnitude of the increase in denaturation temperature induced by disaccharides the binding constants calculated from differential scanning calorimetry are comparable with those extrapolated from titration calorimetry indicating that L-14 interacts with ligands essentially in the folded state.

Factors influencing seasonal and monthly changes in the group size of chital or axis deer in southern India

Chital or axis deer (Axis axis) form fluid groups that change in size temporally and in relation to habitat. Predictions of hypotheses relating animal density, rainfall, habitat structure, and breeding seasonality, to changes in chital group size were assessed simultaneously using multiple regression models of monthly data collected over a 2 yr period in Guindy National Park, in southern India. Over 2,700 detections of chital groups were made during four seasons in three habitats (forest, scrubland and grassland). In scrubland and grassland, chital group size was positively related to animal density, which increased with rainfall. This suggests that in these habitats, chital density increases in relation to food availability, and group sizes increase due to higher encounter rate and fusion of groups. The density of chital in forest was inversely related to rainfall, but positively to the number of fruiting tree species and availability of fallen litter, their forage in this habitat. There was little change in mean group size in the forest, although chital density more than doubled during the dry season and summer. Dispersion of food items or the closed nature of the forest may preclude formation of larger groups. At low densities, group sizes in all three habitats were similar. Group sizes increased with chital density in scrubland and grassland, but more rapidly in the latter—leading to a positive relationship between openness and mean group size at higher densities. It is not clear, however, that this relationship is solely because of the influence of habitat structure. The rutting index (monthly percentage of adult males in hard antler) was positively related to mean group size in forest and scrubland, probably reflecting the increase in group size due to solitary males joining with females during the rut. The fission-fusion system of group formation in chital is thus interactively influenced by several factors. Aspects that need further study, such as interannual variability, are highlighted.

Thermodynamics of Monosaccharide and Disaccharide Binding to Erythrina corallodendron Lectin

Isothermal titration calorimetry measurements of the binding of 2′-fucosyllactose, lactose, N-acetyllactosamine, galactopyranose, 2-acetamido-2-deoxygalactopyranoside, methyl α-N-dansylgalactosaminide (Me-α-DNS-GalN), methyl α-D-galactopyranoside, methyl β-D-galactopyranoside, and fucose to Erythrina corallodendron lectin (ECorL), a dimer with one binding site per subunit, were performed at 283-286 and 297-299 K. The site binding enthalpies, ΔHb, with the exception of Me-α-DNS-GalN, are the same at both temperatures and range from −47.1 ± 1.0 kJ mol−1 for N-acetyllactosamine to −4.4 ± 0.3 kJ mol−1 for fucose, and the site binding constants range from 3.82 ± 0.9 × 105 M−1 for Me-α-DNS-GalN at 283.2 K to 0.46 ± 0.05 × 103 M−1 for fucose at 297.2 K. The binding reactions are mainly enthalpically driven except for fucose and exhibit enthalpy-entropy compensation. The binding enthalpies of the disaccharides are about twice the binding enthalpies of the monosaccharides in contrast to concanavalin A where the binding enthalpies do not double for the disaccharides. Differential scanning calorimetry measurements show that denaturation of the ECorL dimer results in dissociation into its monomer subunits. The binding constants from the increase in denaturation temperature of ECorL in the presence of saccharides are in agreement with values from isothermal titration calorimetry results. The thermal denaturation of ECorL occurs around 333 K, well below the 344-360 K denaturation temperature of other legume lectins of similar size and tertiary structure, undoubtedly due to the difference in its quaternary structure relative to other legume lectins. This is also apparent from the independent unfolding of its two domains.

Cloning and sequencing of winged bean (Psophocarpus tetragonolobus) basic agglutinin (WBA I): presence of second glycosylation site and its implications in quaternary structure

We report cloning of the DNA encoding winged bean basic agglutinin (WBA I). Using oligonucleotide primers corresponding to N- and C-termini of the mature lectin, the complete coding sequence for WBA I could be amplified from genomic DNA. DNA sequence determination by the chain termination method revealed the absence of any intervening sequences in the gene. The DNA deduced amino acid sequence of WBA I displayed some differences with its primary structure established previously by chemical means. Comparison of the sequence of WBA I with that of other legume lectins highlighted several interesting features, including the existence of the largest specificity determining loop which might account for its oligosaccharide-binding specificity and the presence of an additional N-glycosylation site. These data also throw some light on the relationship between the primary structure of the protein and its probable mode of dimerization.

Why do birds participate in mixed-species foraging flocks? A large-scale synthesis

Mixed-species flocks of foraging birds have been documented from terrestrial habitats all over the world and are thought to form for either improved feeding efficiency or better protection from predators. Two kinds of flock participants are recognized: those that join other species ('followers') and are therefore likely to be the recipients of the benefits of flock participation and those that are joined ('leaders'). Through comparative analyses, using a large sample of flocks from around the world, we show that (1) 'followers' tend to be smaller, more insectivorous, and feed in higher strata than matched species that participate in flocks to a lesser extent and (2) 'leaders' tend to be cooperative breeders more often than matched species that are not known to lead flocks. Furthermore, meta-analyses of published results from across the world showed that bird species in terrestrial mixed-species flocks increase foraging rates and reduce vigilance compared to when they are solitary or in conspecific groups. Moreover, the increase in foraging rates is seen only with flock followers and not flock leaders. These findings suggest a role for predation in the evolution of mixed-species flocking. Species that are vulnerable to predation follow species whose vigilance they can exploit. By doing so, they are able to reduce their own vigilance and forage at higher rates. (C) 2009 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.