ИСТИНА

Comparative analysis of homologous proteins in a functionally diverse superfamily is a valuable tool at studying structure-function relationship, but represents a methodological challenge. We have developed an open-access platform available at https://biokinet.belozersky.msu.ru/mustguseal consisting of free on-line methods to study the structure-function relationship in proteins, to select the most promising hot-spots for implementation of novel functions, improvement of stability and evolvability of useful proteins/enzymes, and to design of their selective modulators. The key web-server Mustguseal can automatically collect and align thousands of homologous protein sequences and structures, and four sister web-methods are available for consequent analysis of the collected data: the Zebra web-server to identify variable amino acid residues responsible for functional diversity within a superfamily and to select hotspots for directed evolution or rational design experiments; the pocketZebra web-server to identify and rank binding sites in proteins by their functional significance and to select particular positions in the structure important for selective binding of substrates/inhibitors/effectors; the visualCMAT web-server to select and interpret correlated mutations/co-evolving residues in protein structures; the Yosshi web-server to classify and study disulfide bonds in protein families as well as to select hot-spots for disulfide engineering. Integration of these bioinformatic web-tools provides an out-of-the-box easy-to-use solution, first of its kind, to systematically analyze all the available sequence and structural data related to a protein superfamily, thus promoting the value of bioinformatics for protein engineering and drug discovery. This work was funded by the Russian Foundation for Basic Research [18-29-13060] and carried out using the HPC computing resources at the Lomonosov Moscow State University supported by the project RFMEFI62117X0011.