Several algorithms to identify SSPs/SDPs in a multiple sequence alignment of homologs have been developed ( 3, 7, 9–13). Selection of the specific positions is more complicated. Identification of the conserved positions is relatively straightforward and represents an efficient approach to predict the key functionally important residues from sequence information alone ( 8). Information on both conserved and specific positions can help to understand how the enzyme performs its inherent function, while the latter can also be selected as hotspots for mutation in an attempt to improve the wild-type protein for a particular purpose ( 4–7). The specific positions – also known as ‘subfamily-specific’ ( 1) or ‘family-specific’ ( 2) positions (SSPs, FSPs), ‘specificity-determining residues/sites/positions’ or SDRs/SDSs/SDPs ( 3) etc.-that are conserved only within families/subfamilies, but are different between them, seem to play an important role in functional diversity of homologs. Highly conserved positions are not subjected to evolution and play the functional and/or structural role common for the entire superfamily. Comparative bioinformatic analysis of homologs implementing different properties within the superfamily can help to understand the relationship between protein sequence/structure and its biological function. ![]() ![]() The web-servers are free and open to all users at, no login required.ĭuring evolution of proteins from a common ancestor, one functional property may be preserved, while others may vary as a result of mutations accumulated within a shared 3D-structure, leading to functional diversity. The integration of Zebra2 and Mustguseal web-tools provides the first of its kind out-of-the-box open-access solution to conduct a systematic analysis of evolutionarily related proteins implementing different functions within a shared 3D-structure of the superfamily, determine common and specific patterns of function-associated local structural elements, assist to select hot-spots for rational design and to prepare focused libraries for directed evolution. The results are automatically prioritized and provided at four information levels to facilitate the knowledge-driven expert selection of the most promising positions on-line: as a sequence similarity network interfaces to sequence-based and 3D-structure-based analysis of conservation and variability and accompanied by the detailed annotation of proteins accumulated from the integrated databases with links to the external resources. ![]() The bioinformatic analysis is facilitated by Mustguseal-a companion web-server to automatically collect and superimpose a large representative set of functionally diverse homologs with high structure similarity but low sequence identity to the selected query protein. Explore ways to get support to common device issues at a glance.Zebra2 is a highly automated web-tool to search for subfamily-specific and conserved positions (i.e. the determinants of functional diversity as well as the key catalytic and structural residues) in protein superfamilies. ![]() Zebra’s Self-Service Center is your go-to spot for online tools, resources and support you need to get answers right away.
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