Energy landscape theory, funnels, specificity, and optimal criterion of biomolecular binding

We study the nature of biomolecular binding. We found that in general there exists several thermodynamic phases: a native binding phase, a non-native phase, and a glass or local trapping phase. The quantitative optimal criterion for the binding specificity is found to be the maximization of the ratio of the binding transition temperature versus the trapping transition temperature, or equivalently the ratio of the energy gap of binding between the native state and the average non-native states versus the dispersion or variance of the non-native states. This leads to a funneled binding energy landscape.

Protein Docking by the Underestimation of Free Energy Funnels in the Space of Encounter Complexes

Similarly to protein folding, the association of two proteins is driven by a free energy funnel, determined by favorable interactions in some neighborhood of the native state. We describe a docking method based on stochastic global minimization of funnel-shaped energy functions in the space of rigid body motions (SE(3)) while accounting for flexibility of the interface side chains. The method, called semi-definite programming-based underestimation (SDU), employs a general quadratic function to underestimate a set of local energy minima and uses the resulting underestimator to bias further sampling. While SDU effectively minimizes functions with funnel-shaped basins, its application to docking in the rotational and translational space SE(3) is not straightforward due to the geometry of that space. We introduce a strategy that uses separate independent variables for side-chain optimization, center-to-center distance of the two proteins, and five angular descriptors of the relative orientations of the molecules. The removal of the center-to-center distance turns out to vastly improve the efficiency of the search, because the five-dimensional space now exhibits a well-behaved energy surface suitable for underestimation. This algorithm explores the free energy surface spanned by encounter complexes that correspond to local free energy minima and shows similarity to the model of macromolecular association that proceeds through a series of collisions. Results for standard protein docking benchmarks establish that in this space the free energy landscape is a funnel in a reasonably broad neighborhood of the native state and that the SDU strategy can generate docking predictions with less than 5 � ligand interface Ca root-mean-square deviation while achieving an approximately 20-fold efficiency gain compared to Monte Carlo methods.

Specific and nonspecific collapse in protein folding funnels

Experiments with fast folding proteins are beginning to address the relationship between collapse and folding. We investigate how different scenarios for folding can arise depending on whether the folding and collapse transitions are concurrent or whether a nonspecific collapse precedes folding. Many earlier studies have focused on the limit in which collapse is fast compared to the folding time; in this work we focus on the opposite limit where, at the folding temperature, collapse and folding occur simultaneously. Real proteins exist in both of these limits. The folding mechanism varies substantially in these two regimes. In the regime of concurrent folding and collapse, nonspecific collapse now occurs at a temperature below the folding temperature (but slightly above the glass transition temperature).

Visualization of Protein Folding Funnels in Lattice Models

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Folding funnels and frustration in off-lattice minimalist protein landscapes

A full quantitative understanding of the protein folding problem is now becoming possible with the help of the energy landscape theory and the protein folding funnel concept. Good folding sequences have a landscape that resembles a rough funnel where the energy bias towards the native state is larger than its ruggedness. Such a landscape leads not only to fast folding and stable native conformations but, more importantly, to sequences that are robust to variations in the protein environment and to sequence mutations. In this paper, an off-lattice model of sequences that fold into a β-barrel native structure is used to describe a framework that can quantitatively distinguish good and bad folders. The two sequences analyzed have the same native structure, but one of them is minimally frustrated whereas the other one exhibits a high degree of frustration.

The species composition of true bug assemblages extracted with Berlese funnels (Data to the knowledge on the ground-living Heteroptera of Hungary, No 1)

A rich collection of Heteroptera extracted with Berlese funnel by Dr. I. Loksa between 1953–1974 in Hungary, has been examined. Altogether 157 true bug species have been identified. The great majority of them have been found in very low number, there are only 27 species of which more than 10 adult individuals have been found. Some species considered to be rare or very rare in Hungary have been collected in relatively great number (Ceratocombus coleoptratus, Cryptostemma pusillimum, C. waltli, Acalypta carinata, A. platycheila, Loricula ruficeps, Myrmedobia exilis). The three families, which are more or less rich in species and have the highest ratio of extracted species, were Rhyparochromidae, Tingidae and Nabidae. Out of them, the family Rhyparochromidae has been found to be most diverse and most characteristic at the ground-level. Individuals of the families Tingidae, Hebridae and Rhyparochromidae have been found in greatest number. The occurrence of the lace bug Campylosteira orientalis Horváth, 1881 in Hungary has been verified by a voucher specimen. In respect to the environmental changes through the country, parallel changes have been observed in the zoogeographical distribution of the ground-living bugs.

Spatial and temporal patterns of true bug assemblages extracted with Berlese funnels (Data to the knowledge on the ground-living Heteroptera of Hungary, No. 2)

A rich material of Heteroptera extracted with Berlese funnels by Dr. I. Loksa between 1953–1974 in Hungary, has been examined. Altogether 157 true bug species have been identified. The ground-living heteropteran assemblages collected in different plant communities, substrata, phytogeographical provinces and seasons have been compared with multivariate methods. Because of the unequal number of samples, the objects have been standardized with stochastic simulation. There are several true bug species, which have been collected in almost all of the plant communities. However, characteristic ground-living heteropteran assemblages have been found in numerous Hungarian plant community types. Leaf litter and debris seem to have characteristic bug assemblages. Some differences have also been recognised between the bug fauna of mosses growing on different surfaces. Most of the species have been found in all of the great phytogeographical provinces of Hungary. Most high-dominance species, which have been collected, can be found at the ground-level almost throughout the year. Specimens of many other species have been collected with Berlese funnels in spring, autumn and/or winter. The diversities of the ground-living heteropteran assemblages of the examined objects have also been compared.

Ghost fishing in the pot fishery for blue swimmer crabs Portunus pelagicus in Queensland, Australia

Blue swimmer crabs (Portunus pelagicus) are an economically important crab caught in baited traps throughout the Indo-west Pacific and Mediterranean. In Australia they are traditionally caught using rigid wire traps (approximate to pots) but there has been a recent increase in the use of collapsible pots constructed from polyethylene trawl mesh. Two experiments were conducted in Moreton Bay, Queensland, to determine the ghost fishing potential of lost crab pots on both target and bycatch species and to evaluate the differences between traditional and contemporary pot designs. A lost contemporary, collapsible trawl mesh pot will catch between 3 and 223 R pelagicus per year after the bait has been exhausted, while a traditional wire mesh pot would catch 11-74 crabs peryear. As most fishers now use the collapsible trawl mesh pots, ghost fishing mortality could be as high as 111,811-670,866 crabs per year. Bycatch retention was also higher in contemporary designs. Periods of strong winds appeared to increase the ghost fishing potential of lost pots. The use of escape gaps, larger mesh sizes and construction options that allow for the deterioration of entrance funnels to minimise ghost fishing are recommended to reduce environmental impacts.

Accelerating Waves in Polar Coronal Holes as Seen by EIS and SUMER

We present EIS/Hinode and SUMER/SOHO observations of propagating disturbances detected in coronal lines in inter-plume and plume regions of a polar coronal hole. The observation was carried out on 2007 November 13 as part of the JOP196/HOP045 program. The SUMER spectroscopic observation gives information about fluctuations in radiance and on both resolved (Doppler shift) and unresolved (Doppler width) line-of-sight velocities, whereas EIS 40 `'wide slot images detect fluctuations only in radiance but maximize the probability of overlapping field of view between the two instruments. From distance-time radiance maps, we detect the presence of propagating waves in a polar inter-plume region with a period of 15-20 minutes and a propagation speed increasing from 130 +/- 14 km s(-1) just above the limb to 330 +/- 140 km s(-1) around 160 `' above the limb. These waves can be traced to originate from a bright region of the on-disk part of the coronal hole where the propagation speed is in the range of 25 +/- 1.3 to 38 +/- 4.5 km s(-1), with the same periodicity. These on-disk bright regions can be visualized as the base of the coronal funnels. The adjacent plume region also shows the presence of propagating disturbances with the same range of periodicity but with propagation speeds in the range of 135 +/- 18 to 165 +/- 43 km s(-1) only. A comparison between the distance-time radiance map of the two regions indicates that the waves within the plumes are not observable (may be getting dissipated) far off-limb, whereas this is not the case in the inter-plume region. A correlation analysis was also performed to find out the time delay between the oscillations at several heights in the off-limb region, finding results consistent with those from the analysis of the distance-timemaps. To our knowledge, this result provides first spectroscopic evidence of the acceleration of propagating disturbances in the polar region close to the Sun (within 1.2 R/R-circle dot), which provides clues to the understanding of the origin of these waves. We suggest that the waves are likely either Alfvenic or fast magnetoacoustic in the inter-plume region and slow magnetoacoustic in the plume region. This may lead to the conclusion that inter-plumes are a preferred channel for the acceleration of the fast solar wind.

Escapement of the Cape rock lobster (Jasus lalandii ) through the mesh and entrance of commercial traps

Metal-framed traps covered with polyethylene mesh used in the fishery for the South African Cape rock lobster (Jasus lalandii) incidentally capture large numbers of undersize (<75 mm CL) specimens. Air-exposure, handling, and release procedures affect captured rock lobsters and reduce the productivity of the stock, which is heavily fished. Optimally, traps should retain legalsize rock lobsters and allow sublegal animals to escape before traps are hauled. Escapement, based on lobster morphometric measurements, through meshes of 62 mm, 75 mm, and 100 mm was investigated theoretically under controlled conditions in an aquarium, and during field trials. SELECT models were used to model escapement, wherever appropriate. Size-selectivity curves based on the logistic model fitted the aquarium and field data better than asymmetrical Richards curves. The lobster length at 50% retention (L50) on the escapement curve for 100-mm mesh in the aquarium (75.5 mm CL) approximated the minimum legal size (75 mm CL); however estimates of L50 increased to 77.4 mm in field trials where trapentrances were sealed, and to 82.2 mm where trap-entrances were open. Therfore, rock lobsters that cannot escape through the mesh of sealed field traps do so through the trap entrance of open traps. By contrast, the wider selection range and lower L25 of field, compared to aquarium, trials (SR = 8.2 mm vs. 2.6 mm; L25 =73.4 mm vs. 74.1 mm), indicate that small lobsters that should be able to escape from 100-mm mesh traps do not always do so. Escapement from 62-mm mesh traps with open entrance funnels increased by 40−60% over sealed traps. The findings of this study with a known size distribution, are related to those of a recent indirect (comparative) study for the same species, and implications for trap surveys, commercial catch rates, and ghost fishing are discussed.

The diverse Grania fauna (Clitellata : Enchytraeidae) of the Esperance area, Western Australia, with descriptions of two new species

Seven species of the marine enchytraeid genus Grania Southern, 1913 are described from sediments sampled during the 2003 International Workshop on the Marine Flora and Fauna of Esperance Bay and the Recherche Archipelago, on the southern coast of Western Australia. Two species are new to science, the euryhaline Tasmanian G. dolichura Rota and Erseus, 2000 represents a new record for the state, and the remaining four species were known from other parts of Western Australia. Grania quaerens sp. n. is recognized by having a high chaetal index (= 5 short chaetal foot), small coelomocytes, penial apparati with long whip-like terminal stylets, conspicuous spermathecae with ectally bulbous ducts, and ectally granulated ampullae housing sperm rings in their ental region. Grania sperantia sp. n. is readily distinguishable by the complete lack of lateral chaetae, a multiple-banded pattern of the clitellum, extremely long sperm funnels, and the intrasegmental location of the spermathecal pores. The latter new species and four others in the collection (G. bykane Coates, 1990, G. crassiducta Coates, 1990, G. dolichura, and G. ersei Coates, 1990) are remarkable in possessing the head organ, a sensory structure unique to Grania that was not noted previously in Western Australian species. When considering the whole genus, the geographic pattern of the head organ appears southern-centred: of the 17 species of Grania reported to possess it, as many as 13 inhabit the southern latitudes. The seventh species of the Esperance collection, G. vacivasa Coates and Stacey, 1993, is notable for the kind of items found in its gut and the unusual appearance of its pygidium.

Quantifying intrinsic specificity: A potential complement to affinity in drug screening

We report here the investigation of a novel description of specificity in protein-ligand binding based on energy landscape theory. We define a new term, intrinsic specificity ratio (ISR), which describes the level of discrimination in binding free energies of the native basin for a protein-ligand complex from the weaker binding states of the same ligand. We discuss the relationship between the intrinsic specificity we defined here and the conventional definition of specificity. In a docking study of molecules with the enzyme COX-2, we demonstrate a statistical correspondence between ISR value and geometrical shapes of the small molecules binding to COX-2. We further observe that the known selective (nonselective) inhibitors of COX-2 have higher (lower) ISR values. We suggest that intrinsic specificity ratio may be a useful new criterion and a complement to affinity in drug screening and in searching for potential drug lead compounds.

Quantifying the kinetic paths of flexible biomolecular recognition

Biomolecular recognition often involves large conformational changes, sometimes even local unfolding. The identification of kinetic pathways has become a central issue in understanding the nature of binding. A new approach is proposed here to study the dynamics of this binding-folding process through the establishment of a path-integral framework on the underlying energy landscape. The dominant kinetic paths of binding and folding can be determined and quantified. The significant coupling between the binding and folding of biomolecules often exists in many important cellular processes. In this case, the corresponding kinetic paths of binding are shown to be intimately correlated with those of folding and the dynamics becomes quite cooperative. This implies that binding and folding happen concurrently. When the coupling between binding and folding is weak (strong), the kinetic process usually starts with significant folding (binding) first, with the binding (folding) later proceeding to the end. The kinetic rate can be obtained through the contributions from the dominant paths. The rate is shown to have a bell-shaped dependence on temperature in the concentration-saturated regime consistent with experiment. The changes of the kinetics that occur upon changing the parameters of the underlying binding-folding energy landscape are studied.

Single-molecule dynamics reveals cooperative binding-folding in protein recognition

The study of associations between two biomolecules is the key to understanding molecular function and recognition. Molecular function is often thought to be determined by underlying structures. Here, combining a single-molecule study of protein binding with an energy-landscape-inspired microscopic model, we found strong evidence that biomolecular recognition is determined by flexibilities in addition to structures. Our model is based on coarse-grained molecular dynamics on the residue level with the energy function biased toward the native binding structure ( the Go model). With our model, the underlying free-energy landscape of the binding can be explored. There are two distinct conformational states at the free-energy minimum, one with partial folding of CBD itself and significant interface binding of CBD to Cdc42, and the other with native folding of CBD itself and native interface binding of CBD to Cdc42. This shows that the binding process proceeds with a significant interface binding of CBD with Cdc42 first, without a complete folding of CBD itself, and that binding and folding are then coupled to reach the native binding state.

A comparative study of the effects of metal contamination on collembola in the field and in the laboratory

We examined the species diversity and abundance of Collembola at 32 sampling points along a gradient of metal contamination in a rough grassland site ( Wolverhampton, England), formerly used for the disposal of metal-rich smelting waste. Differences in the concentrations of Cd, Cu, Pb and Zn between the least and most contaminated part of the 35 metre transect were more than one order of magnitude. A gradient of Zn concentrations from 597 to 9080 mug g(-1) dry soil was found. A comparison between field concentrations of the four metals and previous studies on their relative toxicities to Collembola, suggested that Zn is likely to be responsible for any ecotoxicological effects on springtails at this site. Euedaphic ( soil dwelling) Collembola were extracted by placing soil cores into Tullgren funnels and epedaphic ( surface dwelling) species were sampled using pitfall traps. There was no obvious relationship between the total abundance, or a range of commonly used diversity indices, and Zn levels in soils. However, individual species showed considerable differences in abundance. Metal "tolerant'' (e.g., Ceratophysella denticulata) and metal "sensitive'' (e.g., Cryptopygus thermophilus) species could be identified. Epedaphic species appeared to be influenced less by metal contamination than euedaphic species. This difference is probably due to the higher mobility and lower contact with the soil pore water of epedaphic springtails in comparison to euedaphic Collembola. In an experiment exposing the standard test springtail, Folsomia candida, to soils from all 32 sampling points, adult survival and reproduction showed small but significant negative relationships with total Zn concentrations. Nevertheless, juveniles were still produced from eggs laid by females in the most contaminated soils with 9080 mug g(-1) Zn. Folsomia candida is much more sensitive to equivalent concentrations of Zn in the standard OECD soil. Thus, care should be taken in extrapolating the results of laboratory toxicity tests on metals in OECD soil to field soils, in which, the biological availability of contaminants is likely to be lower. Our studies have shown the importance of ecotoxicological effects at the species level. Although there may be no differences in overall abundance, sensitive species that are numerous in contaminated sites, and which may play important roles in decomposition("keystone species'') can be greatly reduced in numbers by pollution.