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Mutations at protein–protein recognition sites alter binding strength by altering the chemical nature of the interacting surfaces. We present a simple surface energy model, parameterized with empirical values, yielding mean energies of ?48 cal mol?1 Å?2 for interactions between hydrophobic surfaces, ?51 to ?80 cal mol?1 Å?2 for surfaces of complementary charge, and 66–83 cal mol?1 Å?2 for electrostatically repelling surfaces, relative to the aqueous phase. This places the mean energy of hydrophobic surface burial at ?24 cal mol?1 Å?2. Despite neglecting configurational entropy and intramolecular changes, the model correlates with empirical binding free energies of a functionally diverse set of rigid‐body interactions (r = 0.66). When used to rerank docking poses, it can place near‐native solutions in the top 10 for 37% of the complexes evaluated, and 82% in the top 100. The method shows that hydrophobic burial is the driving force for protein association, accounting for 50–95% of the cohesive energy. The model is available open‐source from http://life.bsc.es/pid/web/surface_energy/ and via the CCharpPPI web server http://life.bsc.es/pid/ccharppi/ . Proteins 2015; 83:640–650. © 2015 Wiley Periodicals, Inc. 相似文献
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Experimental management control of Opuntia dillenii Haw. and Agave americana L. in Teno Rural Park,Canary Islands
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José R. Arévalo Silvia Fernández‐Lugo Miguel Mellado Tamar de la Concepción 《Plant Species Biology》2015,30(2):137-146
Invasion biology is an important element of global environmental change and represents one of the main threats to biodiversity. American species were introduced to Tenerife after the Spanish conquest during the eighteenth century, as is the case for Agave americana and Opuntia dillenii. The long period of naturalization and adaptation of these species has led them to become two of the most dispersed introduced species of the archipelago. We analyzed several eradication management processes in an area intensively invaded by both O. dillenii and A. americana. Three treatments were randomly applied: mechanical removal, use of herbicide (glyphosate at 10% volume), and mechanical and herbicide applied together. Both the effectiveness of the treatments to remove the target exotic species biovolume and the impact of the eradication methods on species richness and species composition of the area were analyzed. We found that the treatments had an impact on species composition but not on species richness. Species composition was mainly affected by mechanical treatment. The effect caused by the mechanical removal of the exotic target species in species composition is minor after 4 years, and is related to a higher dominance of shrub species typical of coastal shrubland and of annual or pioneer species. The control of O. dillenii and A. americana is evident from insignificant recovery 4 years after treatment application. A mechanical and herbicide treatment together, allowed not only the immediate removal of large individuals but also the herbicidal control of smaller ones. 相似文献
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Agnieszka?Sitarska Lukasz?Skora Julia?Klopp Susan?Roest César?Fernández Binesh?Shrestha Alvar?D.?GossertEmail authorView authors OrcID profile 《Journal of biomolecular NMR》2015,62(2):191-197
For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80 % can be achieved for 15N and 13C with yields comparable to expression in full media. For 2H,15N and 2H,13C,15N labeling, incorporation is only slightly lower with 75 and 73 %, respectively, and yields are typically twofold reduced. The media were optimized for isotope incorporation, reproducibility, simplicity and cost. High isotope incorporation levels for all labeling patterns are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The final formulation consists of just five commercially available components, at costs 12-fold lower than labeling media from vendors. The approach was applied to several cytosolic and secreted target proteins. 相似文献
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Structural studies of the C‐terminal 19‐peptide of serum amyloid A and its Pro→Ala variants interacting with human cystatin C
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Martyna Maszota Natalia Karska Marta Spodzieja Jerzy Ciarkowski Aleksandra S. Koodziejczyk Sylwia Rodziewicz‐Motowido Paulina Czaplewska 《Journal of molecular recognition : JMR》2015,28(7):413-426
Serum amyloid A (SAA) is a multifunctional acute‐phase protein whose concentration in serum increases markedly following a number of chronic inflammatory and neoplastic diseases. Prolonged high SAA level may give rise to reactive systemic amyloid A (AA) amyloidosis, where the N‐terminal segment of SAA is deposited as amyloid fibrils. Besides, recently, well‐documented association of SAA with high‐density lipoprotein or glycosaminoglycans, in particular heparin/heparin sulfate (HS), and specific interaction between SAA and human cystatin C (hCC), the ubiquitous inhibitor of cysteine proteases, was proved. Using a combination of selective proteolytic excision and high‐resolution mass spectrometry, a hCC binding site in the SAA sequence was determined as SAA(86–104). The role of this SAA C‐terminal fragment as a ligand‐binding locus is still not clear. It was postulated important in native SAA folding and in pathogenesis of AA amyloidosis. In the search of conformational details of this SAA fragment, we did its structure and affinity studies, including its selected double/triple Pro→Ala variants. Our results clearly show that the SAA(86–104) 19‐peptide has rather unordered structure with bends in its C‐terminal part, which is consistent with the previous results relating to the whole protein. The results of affinity chromatography, fluorescent ELISA‐like test, CD and NMR studies point to an importance of proline residues on structure of SAA(86–104). Conformational details of SAA fragment, responsible for hCC binding, may help to understand the objective of hCC–SAA complex formation and its importance for pathogenesis of reactive amyloid A amyloidosis. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献