Analysis of geometrical and topological attitude for proteinprotein interaction

Protein-protein interaction takes usually place on an extended area of the external molecules surfaces that are morphologically fitting. Geometric and topological congruence (i.e. concavity and convexity correspondences) is required to support the neighboring interaction of surface patches belonging to the two protein molecules. It is therefore important to adopt representations and data structures that can facilitate the analysis and the implementation of techniques for the evaluation of geometric and topological properties on extended surfaces. These areas of activity are usually roughly “planar” but with local concavity and complexity that must match each other for interacting. To this purpose we are suggesting a solution different from the one of ligand-protein interaction in which are involved a pocket and a small molecule. The solution here suggested is based on the concavity tree representation. Starting from the convex hull of the protein molecule a recursive process leads to a series of concavity and meta-concavity that allows reaching the detail level required. The consequence of the recursive process is obviously a hierarchical data structure (a tree) which at each level supports a complete description of a surface. Each node of the tree contains an array of features that support the geometrical, topological and biochemical properties of the correspondent surface patch

Components of the E. coli envelope are affected by and can react to protein over-production in the cytoplasm.

BACKGROUND: Protein over-expression in bacteria is still the easiest, cheapest and therefore preferred way to obtain large amounts of proteins for industrial and laboratory scale preparations. Several studies emphasized the importance of understanding cellular and molecular mechanisms triggered by protein over-production in order to obtain higher yield and better quality of the recombinant product. Almost every step leading to a fully functional polypeptide has been investigated, from mRNA stability to the role of molecular chaperones, from aggregation to bottlenecks in the secretory pathway. In this context, we focused on the still poorly addressed relationship between protein production in the cytoplasm and the bacterial envelope, an active and reactive cell compartment that controls interactions with the environment and several major cellular processes. Results available to date show that the accumulation of foreign proteins in the cytoplasm induces changes in the membrane lipids and in the levels of mRNAs for some membrane proteins. However, a direct connection between membrane protein expression levels and soluble/aggregated protein accumulation in the cytoplasm has never been reported. RESULTS: By the use of a combined physiological and proteomic approach, we investigated the effects on the cell membrane of E. coli of the overexpression of two recombinant proteins, the B. cepacia lipase (BCL) and the green fluorescent protein (GFP). Both polypeptides are expressed in the cytoplasm at similar levels but GFP is fully soluble whereas inactive BCL accumulates in inclusion bodies.Growth and viability of the transformed cells were tested in the presence of different drugs. We found that chloramphenycol preferentially inhibited the strain over-producing GFP while SDS was more effective when BCL inclusion bodies accumulated in the cytoplasm. In contrast, both proteins induced a similar response in the membrane proteome, i.e. increased levels of LamB, OmpF, OmpA and TolC. Under all tested conditions, the lipopolysaccharide was not affected, suggesting that a specific rather than a generalized rearrangement of the envelope was induced. CONCLUSION: Taking together physiological and biochemical evidence, our work indicates that the E. coli envelope can sense protein over-expression in the cytoplasm and react by modulating the abundance of some membrane proteins, with possible consequences on the membrane traffic of small solutes, i.e. nutrients, drugs and metabolites. Such a response seems to be independent on the nature of the protein being over-expressed. On the other hand both our data reported herein and previous results indicate that membrane lipids may act as a second stress sensor responsive to the aggregation state of the recombinant protein and further contribute to changes in cellular exchanges with the environment.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Components of the E. coli envelope are affected by and can react to protein over-production in the cytoplasm

<p>Abstract</p> <p>Background</p> <p>Protein over-expression in bacteria is still the easiest, cheapest and therefore preferred way to obtain large amounts of proteins for industrial and laboratory scale preparations. Several studies emphasized the importance of understanding cellular and molecular mechanisms triggered by protein over-production in order to obtain higher yield and better quality of the recombinant product. Almost every step leading to a fully functional polypeptide has been investigated, from mRNA stability to the role of molecular chaperones, from aggregation to bottlenecks in the secretory pathway. In this context, we focused on the still poorly addressed relationship between protein production in the cytoplasm and the bacterial envelope, an active and reactive cell compartment that controls interactions with the environment and several major cellular processes. Results available to date show that the accumulation of foreign proteins in the cytoplasm induces changes in the membrane lipids and in the levels of mRNAs for some membrane proteins. However, a direct connection between membrane protein expression levels and soluble/aggregated protein accumulation in the cytoplasm has never been reported.</p> <p>Results</p> <p>By the use of a combined physiological and proteomic approach, we investigated the effects on the cell membrane of <it>E. coli </it>of the overexpression of two recombinant proteins, the <it>B. cepacia </it>lipase (BCL) and the green fluorescent protein (GFP). Both polypeptides are expressed in the cytoplasm at similar levels but GFP is fully soluble whereas inactive BCL accumulates in inclusion bodies.</p> <p>Growth and viability of the transformed cells were tested in the presence of different drugs. We found that chloramphenycol preferentially inhibited the strain over-producing GFP while SDS was more effective when BCL inclusion bodies accumulated in the cytoplasm. In contrast, both proteins induced a similar response in the membrane proteome, i.e. increased levels of LamB, OmpF, OmpA and TolC. Under all tested conditions, the lipopolysaccharide was not affected, suggesting that a specific rather than a generalized rearrangement of the envelope was induced.</p> <p>Conclusion</p> <p>Taking together physiological and biochemical evidence, our work indicates that the <it>E. coli </it>envelope can sense protein over-expression in the cytoplasm and react by modulating the abundance of some membrane proteins, with possible consequences on the membrane traffic of small solutes, i.e. nutrients, drugs and metabolites. Such a response seems to be independent on the nature of the protein being over-expressed. On the other hand both our data reported herein and previous results indicate that membrane lipids may act as a second stress sensor responsive to the aggregation state of the recombinant protein and further contribute to changes in cellular exchanges with the environment.</p

Nanoindentation of Au nanoparticles – A combined experimental/computational multiscale study

The idea of dimensionality and size effect the strength of metallic specimen as their typical size is pushed into the sub-micrometer scale is well established. The importance of the shape at the nanoscale was demonstrated on Au thin-films and nanoparticles in nanoindentation experiments. It was shown that nanoparticles are substantially softer than thin-films of the same height and the smallest nanoparticles are softer than the largest ones [1]. We propose that the size effect arises from the interaction between the lateral free surfaces on the plastic zone. However, experiments alone cannot provide the understanding on the governing microstructural dislocation mechanisms and we demonstrate here a combined experimental/computational study, by developing a multiscale frame to study nanoindentation of nanoparticles from the atomic- to the macro-scale. Please click Additional Files below to see the full abstract