Enzyme function and its evolution

With rapid increases over recent years in the determination of protein sequence and structure, alongside knowledge of thousands of enzyme functions and hundreds of chemical mechanisms, it is now possible to combine breadth and depth in our understanding of enzyme evolution. Phylogenetics continues to move forward, though determining correct evolutionary family trees is not trivial. Protein function prediction has spawned a variety of promising methods that offer the prospect of identifying enzymes across the whole range of chemical functions and over numerous species. This knowledge is essential to understand antibiotic resistance, as well as in protein re-engineering and de novo enzyme design.PostprintPeer reviewe

Nano-topography and functionalization with the synthetic peptoid GN2-Npm9 as a strategy for antibacterial and biocompatible titanium implants

In recent years, antimicrobial peptides (AMPs) have attracted great interest in scientific research, especially for biomedical applications such as drug delivery and orthopedic applications. Since they are readily degradable in the physiological environment, scientific research has recently been trying to make AMPs more stable. Peptoids are synthetic N-substituted glycine oligomers that mimic the structure of peptides. They have a structure that does not allow proteolytic degradation, which makes them more stable while maintaining microbial activity. This structure also brings many advantages to the molecule, such as greater diversity and specificity, making it more suitable for biological applications. For the first time, a synthesized peptoid (GN2-Npm9) was used to functionalize a nanometric chemically pre-treated (CT) titanium surface for bone-contact implant applications. A preliminary characterization of the functionalized surfaces was performed using the contact angle measurements and zeta potential titration curves. These preliminary analyses confirmed the presence of the peptoid and its adsorption on CT. The functionalized surface had a hydrophilic behaviour (contact angle = 30°) but the hydrophobic tryptophan-like residues were also exposed. An electrostatic interaction between the lysine residue of GN2-Npm9 and the surface allowed a chemisorption mechanism. The biological characterization of the CT_GN2-Nmp9 surfaces demonstrated the ability to prevent surface colonization and biofilm formation by the pathogens Escherichia coli and Staphylococcus epidermidis thus showing a broad-range activity. The cytocompatibility was confirmed by human mesenchymal stem cells. Finally, a bacteria-cells co-culture model was applied to demonstrate the selective bioactivity of the CT_GN2-Nmp9 surface that was able to preserve colonizing cells adhered to the device surface from bacterial infection

An assessment of catalytic residue 3D ensembles for the prediction of enzyme function

Background The central element of each enzyme is the catalytic site, which commonly catalyzes a single biochemical reaction with high specificity. It was unclear to us how often sites that catalyze the same or highly similar reactions evolved on different, i. e. non-homologous protein folds and how similar their 3D poses are. Both similarities are key criteria for assessing the usability of pose comparison for function prediction. Results We have analyzed the SCOP database on the superfamily level in order to estimate the number of non-homologous enzymes possessing the same function according to their EC number. 89 % of the 873 substrate-specific functions (four digit EC number) assigned to mono-functional, single-domain enzymes were only found in one superfamily. For a reaction-specific grouping (three digit EC number), this value dropped to 35 %, indicating that in approximately 65 % of all enzymes the same function evolved in two or more non-homologous proteins. For these isofunctional enzymes, structural similarity of the catalytic sites may help to predict function, because neither high sequence similarity nor identical folds are required for a comparison. To assess the specificity of catalytic 3D poses, we compiled the redundancy-free set ENZ_SITES, which comprises 695 sites, whose composition and function are well-defined. We compared their poses with the help of the program Superpose3D and determined classification performance. If the sites were from different superfamilies, the number of true and false positive predictions was similarly high, both for a coarse and a detailed grouping of enzyme function. Moreover, classification performance did not improve drastically, if we additionally used homologous sites to predict function. Conclusions For a large number of enzymatic functions, dissimilar sites evolved that catalyze the same reaction and it is the individual substrate that determines the arrangement of the catalytic site and its local environment. These substrate-specific requirements turn the comparison of catalytic residues into a weak classifier for the prediction of enzyme function

Additional file 1: of An assessment of catalytic residue 3D ensembles for the prediction of enzyme function

Composition of data set ENZ_SITES. (PDF 84 kb