Statistical analysis and prediction of protein-protein interfaces

Predicting protein-protein interfaces from a three-dimensional structure is a key task of computational structural proteomics. In contrast to geometrically distinct small molecule binding sites, protein-protein interface are notoriously difficult to predict. We generated a large nonredundant data set of 1494 true protein-protein interfaces using biological symmetry annotation where necessary. The data set was carefully analyzed and a Support Vector Machine was trained on a combination of a new robust evolutionary conservation signal with the local surface properties to predict protein-protein interfaces. Fivefold cross validation verifies the high sensitivity and selectivity of the model. As much as 97% of the predicted patches had an overlap with the true interface patch while only 22% of the surface residues were included in an average predicted patch. The model allowed the identification of potential new interfaces and the correction of mislabeled oligomeric states.     

Hydration of protein-protein interfaces

We present an analysis of the water molecules immobilized at the protein-protein interfaces of 115 homodimeric proteins and 46 protein-protein complexes, and compare them with 173 large crystal packing interfaces representing nonspecific interactions. With an average of 15 waters per 1000 A2 of interface area, the crystal packing interfaces are more hydrated than the specific interfaces of homodimers and complexes, which have 10-11 waters per 1000 A2, reflecting the more hydrophilic composition of crystal packing interfaces. Very different patterns of hydration are observed: Water molecules may form a ring around interfaces that remain "dry," or they may permeate "wet" interfaces. A majority of the specific interfaces are dry and most of the crystal packing interfaces are wet, but counterexamples exist in both categories. Water molecules at interfaces form hydrogen bonds with protein groups, with a preference for the main-chain carbonyl and the charged side-chains of Glu, Asp, and Arg. These interactions are essentially the same in specific and nonspecific interfaces, and very similar to those observed elsewhere on the protein surface. Water-mediated polar interactions are as abundant at the interfaces as direct protein-protein hydrogen bonds, and they may contribute to the stability of the assembly.     

New measures for estimating surface complementarity and packing at protein-protein interfaces

A number of methods exist that use different approaches to assess geometric properties like the surface complementarity and atom packing at the protein-protein interface. We have developed two new and conceptually different measures using the Delaunay tessellation and interface slice selection to compute the surface complementarity and atom packing at the protein-protein interface in a straightforward manner. Our measures show a strong correlation among themselves and with other existing measures, and can be calculated in a highly time-efficient manner. The measures are discriminative for evaluating biological, as well as non-biological protein-protein contacts, especially from large protein complexes and large-scale structural studies (http://pallab.serc.iisc.ernet.in/nip_nsc).     

Prediction of quaternary structure by analysis of hot spot residues in protein-protein interfaces: the case of anthranilate phosphoribosyltransferases

It is an important goal of computational biology to correctly predict the association state of a protein based on its amino acid sequence and the structures of known homologues. We have pursued this goal on the example of anthranilate phosphoribosyltransferase (AnPRT), an enzyme that is involved in the biosynthesis of the amino acid tryptophan. Firstly, known crystal structures of naturally occurring homodimeric AnPRTs were analyzed using the Protein Interfaces, Surfaces, and Assemblies (PISA) service of the European Bioinformatics Institute (EBI). This led to the identification of two hydrophobic “hot spot” amino acids in the protein-protein interface that were predicted to be essential for self-association. Next, in a comprehensive multiple sequence alignment (MSA), naturally occurring AnPRT variants with hydrophilic or charged amino acids in place of hydrophobic residues in the two hot spot positions were identified. Representative variants were characterized in terms of thermal stability, enzymatic activity, and quaternary structure. We found that AnPRT variants with charged residues in both hot spot positions exist exclusively as monomers in solution. Variants with hydrophilic amino acids in one hot spot position occur in both forms, monomer and dimer. The results of the present study provide a detailed characterization of the determinants of the AnPRT monomer-dimer equilibrium and show that analysis of hot spots in combination with MSAs can be a valuable tool in prediction of protein quaternary structures.     

Comprehensive statistical analysis of residues interaction specificity at protein-protein interfaces

We calculated interchain contacts on the atomic level for nonredundant set of 4602 protein-protein interfaces using an unbiased Voronoi-Delaune tessellation method, and made 20x20 residue contact matrixes both for homodimers and heterocomplexes. The area of contacts and the distance distribution for these contacts were calculated on both the residue and the atomic levels. We analyzed residue area distribution and showed the existence of two types of interresidue contacts: stochastic and specific. We also derived formulas describing the distribution of contact area for stochastic and specific interactions in parametric form. Maximum pairing preference index was found for Cys-Cys contacts and for oppositely charged interactions. A significant difference in residue contacts was observed between homodimers and heterocomplexes. Interfaces in homodimers were enriched with contacts between residues of the same type due to the effects of structure symmetry.     

Statistical analysis of atomic contacts at RNA-protein interfaces

Forty-five crystals of complexes between proteins and RNA molecules from the Protein Data Bank have been statistically surveyed for the number of contacts between RNA components (phosphate, ribose and the four bases) and amino acid side chains. Three groups of complexes were defined: the tRNA synthetases; the ribosomal complexes; and a third group containing a variety of complexes. The types of atomic contacts were a priori classified into ionic, neutral H-bond, C-H...O H-bond, or van der Waals interaction. All the contacts were organized into a relational database which allows for statistical analysis. The main conclusions are the following: (i) in all three groups of complexes, the most preferred amino acids (Arg, Asn, Ser, Lys) and the less preferred ones (Ala, Ile, Leu, Val) are the same; Trp and Cys are rarely observed (respectively 15 and 5 amino acids in the ensemble of interfaces); (ii) of the total number of amino acids located at the interfaces 22% are hydrophobic, 40% charged (positive 32%, negative 8%), 30% polar and 8% are Gly; (iii) in ribosomal complexes, phosphate is preferred over ribose, which is preferred over the bases, but there is no significant preference in the other two groups; (iv) there is no significant prevalence of a base type at protein-RNA interfaces, but specifically Arg and Lys display a preference for phosphate over ribose and bases; Pro and Asn prefer bases over ribose and phosphate; Met, Phe and Tyr prefer ribose over phosphate and bases. Further, Ile, Pro, Ser prefer A over the others; Leu prefers C; Asp and Gly prefer G; and Asn prefers U. Considering the contact types, the following conclusions could be drawn: (i) 23% of the contacts are via potential H-bonds (including CH...O H-bonds and ionic interactions), 72% belong to van der Waals interactions and 5% are considered as short contacts; (ii) of all potential H-bonds, 54% are standard, 33% are of the C-H...O type and 13% are ionic; (iii) the Watson-Crick sites of G, O6(G) and principally N2(G) and the hydroxyl group O2' is more often involved in H-bonds than expected; the protein main chain is involved in 32% and the side chains in 68% of the H-bonds; considering the neutral and ionic H-bonds, the following couples are more frequent than expected-base A-Ser, base G-Asp/Glu, base U-Asn. The RNA CH groups interact preferentially with oxygen atoms (62% on the main chain and 19% on the side chains); (iv) the bases are involved in 38% of all H-bonds and more than 26% of the H-bonds have the H donor group on the RNA; (v) the atom O2' is involved in 21% of all H-bonds, a number greater than expected; (vi) amino acids less frequently in direct contact with RNA components interact frequently via their main chain atoms through water molecules with RNA atoms; in contrast, those frequently observed in direct contact, except Ser, use instead their side chain atoms for water bridging interactions.     

Hydrophobic patches on protein subunit interfaces: Characteristics and prediction

Hydrophobic patches, defined as clusters of neighboring apolar atoms deemed accessible on a given protein surface, have been investigated on protein subunit interfaces. The data were taken from known tertiary structures of multimeric protein complexes. Amino acid composition and preference, patch size distribution, and patch contact complementarity across associating subunits were examined and compared with hydrophobic patches found on the solvent-accessible surface of the multimeric complexes. The largest or second largest patch on the accessible surface of the entire subunit was involved in multimeric interfaces in 90% of the cases. These results should prove useful for subunit design and engineering as well as for prediction of subunit interface regions. Proteins 28:333–343, 1997. © 1997 Wiley-Liss, Inc.     

Architectures and functional coverage of protein-protein interfaces

The diverse range of cellular functions is performed by a limited number of protein folds existing in nature. One may similarly expect that cellular functional diversity would be covered by a limited number of protein-protein interface architectures. Here, we present 8205 interface clusters, each representing a unique interface architecture. This data set of protein-protein interfaces is analyzed and compared with older data sets. We observe that the number of both biological and crystal interfaces increases significantly compared to the number of Protein Data Bank entries. Furthermore, we find that the number of distinct interface architectures grows at a much faster rate than the number of folds and is yet to level off. We further analyze the growth trend of the functional coverage by constructing functional interaction networks from interfaces. The functional coverage is also found to steadily increase. Interestingly, we also observe that despite the diversity of interface architectures, some are more favorable and frequently used, and of particular interest, are the ones that are also preferred in single chains.     

A dissection of specific and non-specific protein-protein interfaces

We compare the geometric and physical-chemical properties of interfaces involved in specific and non-specific protein-protein interactions in crystal structures reported in the Protein Data Bank. Specific interactions are illustrated by 70 protein-protein complexes and by subunit contacts in 122 homodimeric proteins; non-specific interactions are illustrated by 188 pairs of monomeric proteins making crystal-packing contacts selected to bury more than 800 A2 of protein surface. A majority of these pairs have 2-fold symmetry and form "crystal dimers" that cannot be distinguished from real dimers on the basis of the interface size or symmetry. The chemical and amino acid compositions of the large crystal-packing interfaces resemble the protein solvent-accessible surface. These interfaces are less hydrophobic than in homodimers and contain much fewer fully buried atoms. We develop a residue propensity score and a hydrophobic interaction score to assess preferences seen in the chemical and amino acid compositions of the different types of interfaces, and we derive indexes to evaluate the atomic packing, which we find to be less compact at non-specific than at specific interfaces. We test the capacity of these parameters to identify homodimeric proteins in crystal structures, and show that a simple combination of the non-polar interface area and the fraction of buried interface atoms assigns the quaternary structure of 88% of the homodimers and 77% of the monomers in our data set correctly. These success rates increase to 93-95% when the residue propensity score of the interfaces is taken into consideration.     

Biological units and their effect upon the properties and prediction of protein-protein interactions

Structural data as collated in the Protein Data Bank (PDB) have been widely applied in the study and prediction of protein-protein interactions. However, since the basic PDB Entries contain only the contents of the asymmetric unit rather than the biological unit, some key interactions may be missed by analysing only the PDB Entry. A total of 69,054 SCOP (Structural Classification of Proteins) domains were examined systematically to identify the number of additional novel interacting domain pairs and interfaces found by considering the biological unit as stored in the PQS (Protein Quaternary Structure) database. The PQS data adds 25,965 interacting domain pairs to those seen in the PDB Entries to give a total of 61,783 redundant interacting domain pairs. Redundancy filtering at the level of the SCOP family shows PQS to increase the number of novel interacting domain-family pairs by 302 (13.3%) from 2277, but only 16/302 (1.4%) of the interacting domain pairs have the two domains in different SCOP families. This suggests the biological units add little to the elucidation of novel biological interaction networks. However, when the orientation of the domain pairs is considered, the PQS data increases the number of novel domain-domain interfaces observed by 1455 (34.5%) to give 5677 non-redundant domain-domain interfaces. In all, 162/1455 novel domain-domain interfaces are between domains from different families, an increase of 8.9% over the PDB Entries. Overall, the PQS biological units provide a rich source of novel domain-domain interfaces that are not seen in the studied PDB Entries, and so PQS domain-domain interaction data should be exploited wherever possible in the analysis and prediction of protein-protein interactions.     

Improvement of the synthesis and pharmacokinetic properties of chromenotriazolopyrimidine MDM2-p53 protein-protein inhibitors

Human murine double minute 2 (MDM2) is a negative regulator of p53, which plays an important role in cell cycle and apoptosis. We report several optimizations to the synthesis of the chromenotriazolopyrimidine series of MDM2-p53 protein-protein interaction inhibitors. Additionally, the in vitro and in vivo stability, pharmacokinetic properties and solubility were improved through N-substitution.     

Geometrical and electro-static determinants of protein-protein interactions

Protein-protein interactions (PPI) are pivotal to the numerous processes in the cell. Therefore, it is of interest to document the analysis of these interactions in terms of binding sites, topology of the interacting structures and physiochemical properties of interacting interfaces and the of forces interactions. The interaction interface of obligatory protein-protein complexes differs from that of the transient interactions. We have created a large database of protein-protein interactions containing over100 thousand interfaces. The structural redundancy was eliminated to obtain a non-redundant database of over 2,265 interaction interfaces. Therefore, it is of interest to document the analysis of these interactions in terms of binding sites, topology of the interacting structures and physiochemical properties of interacting interfaces and the offorces interactions. The residue interaction propensity and all of the rest of the parametric scores converged to a statistical indistinguishable common sub-range and followed the similar distribution trends for all three classes of sequence-based classifications PPInS. This indicates that the principles of molecular recognition are dependent on the preciseness of the fit in the interaction interfaces. Thus, it reinforces the importance of geometrical and electrostatic complementarity as the main determinants for PPIs.     

A multivariate study of the relationship between the genetic code and the physical-chemical properties of amino acids

Summary The 20 naturally occurring amino acids are characterized by 20 variables: pK_{NH 2}, pK_COOH, pI, molecular weight, substituent van der Waals volume, seven¹H and¹³C nuclear magnetic resonance shift variables, and eight hydrophobicity-hydrophilicity scales. The 20-dimensional data set is reduced to a few new dimensions by principal components analysis. The three first principal components reveal relationships between the properties of the amino acids and the genetic code. Thus the amino acids coded for by adenosine (A), uracil (U), or cytosine (C) in their second codon position (corresponding to U, A, or G in the second anticodon position) are grouped in these components. No grouping was detected for the amino acids coded for by guanine (G) in the second codon position (corresponding to C in the second anticodon position). The results show that a relationship exists between the physical-chemical properties of the amino acids and which of the A (U), U (A), or C (G) nucleotide is used in the second codon (anticodon) position. The amino acids coded for by G (C) in the second codon (anticodon) position do not participate in this relationship.     

Studies of protein-protein interfaces: a statistical analysis of the hydrophobic effect.

Data sets of 362 structurally nonredundant protein-protein interfaces and of 57 symmetry-related oligomeric interfaces have been used to explore whether the hydrophobic effect that guides protein folding is also the main driving force for protein-protein associations. The buried nonpolar surface area has been used to measure the hydrophobic effect. Our analysis indicates that, although the hydrophobic effect plays a dominant role in protein-protein binding, it is not as strong as that observed in the interior of protein monomers. Comparison of interiors of the monomers with those of the interfaces reveals that, in general, the hydrophobic amino acids are more frequent in the interior of the monomers than in the interior of the protein-protein interfaces. On the other hand, a higher proportion of charged and polar residues are buried at the interfaces, suggesting that hydrogen bonds and ion pairs contribute more to the stability of protein binding than to that of protein folding. Moreover, comparison of the interior of the interfaces to protein surfaces indicates that the interfaces are poorer in polar/charged than the surfaces and are richer in hydrophobic residues. The interior of the interfaces appears to constitute a compromise between the stabilization contributed by the hydrophobic effect on the one hand and avoiding patches on the protein surfaces that are too hydrophobic on the other. Such patches would be unfavorable for the unassociated monomers in solution. We conclude that, although the types of interactions are similar between protein-protein interfaces and single-chain proteins overall, the contribution of the hydrophobic effect to protein-protein associations is not as strong as to protein folding. This implies that packing patterns and interatom, or interresidue, pairwise potential functions, derived from monomers, are not ideally suited to predicting and assessing ligand associations or design. These would perform adequately only in cases where the hydrophobic effect at the binding site is substantial.     

菜园土壤微生物生态特征与土壤理化性质的关系

对广州白云区64个菜园土壤样本的研究表明,微生物碳与土壤全氮、碱解氮、有效钾、阳离子交换量和有机质,微生物氮与土壤全氮、全磷、阳离子交换量及有机质,土壤基础呼吸与土壤全氮、碱解氮、全钾、阳离子交换量及有机质,AWCD值与全氮及有机质含量,以及Shannon多样性指数与全氮和阳离子交换量均呈显著正相关关系.较低的碱解氮含量使土壤微生物碳、土壤基础呼吸和呼吸商的值升高,过高的碱解氮则使呼吸商下降;过高的土壤有效磷降低微生物碳、微生物氮和微生物呼吸商.有效磷/碱解氮比值过高降低了土壤微生物碳、微生物氮、微生物碳氮比及土壤基础呼吸.土壤微生物生态特征与土壤理化性质关系密切,有效养分过高及养分比例不适当对土壤微生物均存在不利影响.     

A combinatorial score to distinguish biological and nonbiological protein-protein interfaces

With the large amount of protein-protein complex structural data available, to understand the key features governing the specificity of protein-protein recognition and to define a suitable scoring function for protein-protein interaction predictions, we have analyzed the protein interfaces from geometric and energetic points of view. Atom-based potential of mean force (PMFScore), packing density, contact size, and geometric complementarity are calculated for crystal contacts in 74 homodimers and 91 monomers, which include real biological interactions in dimers and nonbiological contacts in monomers and dimers. Simple cutoffs were developed for single and combinatorial parameters to distinguish biological and nonbiological contacts. The results show that PMFScore is a better discriminator between biological and nonbiological interfaces comparable in size. The combination of PMFScore and contact size is the most powerful pairwise discriminator. A combinatorial score (CFPScore) based on the four parameters was developed, which gives the success rate of the homodimer discrimination of 96.6% and error rate of the monomer discrimination of 6.0% and 19.8% according to Valdar's and our definition, respectively. Compared with other statistical learning models, the cutoffs for the four parameters and their combinations are directly based on physical models, simple, and can be easily applied to protein-protein interface analysis and docking studies.     

蛋白质表面氨基酸特性研究进展

分布在蛋白质分子表面的暴露于溶剂的氨基酸所具有的一些特性对蛋白质的折叠和聚合过程、蛋白质一蛋白质相互作用以及蛋白质的功能具有重要影响。本文分析了蛋白质表面氨基酸在疏水性、保守性、静电势及结构方面的一些特性,阐述了近年来国际上利用这些特性对蛋白质一蛋白质相互作用界面进行预测的方法,最后介绍了几款预测蛋白质表面氨基酸的软件。     

蛋白质表面氨基酸特性研究进展

分布在蛋白质分子表面的暴露于溶剂的氨基酸所具有的一些特性对蛋白质的折叠和聚合过程、蛋白质-蛋白质相互作用以及蛋白质的功能具有重要影响。本文分析了蛋白质表面氨基酸在疏水性、保守性、静电势及结构方面的一些特性,阐述了近年来国际上利用这些特性对蛋白质-蛋白质相互作用界面进行预测的方法,最后介绍了几款预测蛋白质表面氨基酸的软件。     

Soil properties along altitudinal gradient in Himalayan temperate forest of Garhwal region

The aim of the present study was to analyse the soil properties in different seasons at varying altitudes. The study was carried out in Dhanaulti forest, falls under temperate region of Garhwal Himalaya in Uttarakhand State, India. Physical properties and chemical properties of the soil were estimated using all standard procedures. In the present study, sand particles were reported highest (77.21%) in rainy season followed by in summer (70.17%) and winter (63.15%) seasons. The silt and clay particles also followed similar trend as sand which reduced in order of rainy > summer > winter seasons. The water holding capacity of soil ranged from 62.13 to 67.70%. The majority of soils were dark brown to dark yellowish brown in colour, which is considered having higher potential of water holding capacity. The values of nitrogen ranged between 0.01 to 0.012% (upper altitude), 0.009 to 0.011% (middle altitude) and 0.007 to 0.011% (lower altitude). The effects of altitudes and seasons in nitrogen show significant variation. Potassium ranged between 102.29 and 206.22 kg ha^? 1. The combined effect of season and soil-depth also showed significant variation in level of potassium. The soil organic carbon values were between 0.14 and 0.19% and pH values ranged between 6.33 and 6.75 which was slightly acidic in nature.     

Studies on molecular properties prediction,antitubercular and antimicrobial activities of novel quinoline based pyrimidine motifs

In the present study, a series of 3-((6-(2,6-dichloroquinolin-3-yl)-4-aryl-1,6-dihydro-pyrimidin-2-yl)thio)propanenitriles 5a–o were synthesized and subjected to molecular properties prediction and drug-likeness model score by Molinspiration property calculation toolkit and MolSoft software, respectively. Compound 5m (4-OCH₃) was found to be maximum drug-likeness model score (0.42). Among the screened compounds, 5m showed the most promising antitubercular activity with MIC of 0.20 μg/mL, while compounds 5g, 5k and 5m displayed broad spectrum antibacterial activity against all the bacterial strains. Moreover, compound 5k was found to be the most potent antifungal agent. Further, the results of preliminary MTT cytotoxicity studies on HeLa cells suggested that potent antimicrobial activity of 5g, 5k and 5m was escorted by low cytotoxicity.