<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases"^^ . "The current goal of research in the field of synthetic chemistry is the design and production of peptide \r\nand protein structures that function as biological synthesis machines.[1] Initially, the de novo design of \r\npeptide catalysts focused on α-helical structural motifs, such as the coiled coil.[2,3] Given the relatively \r\nrigid and self-assembling nature of these scaffolds[4,5], scientific efforts are concentrated on the design\r\nof smaller single-chain β-sheet motifs, such as the WW domain.[6–9] Due to its properties as a small, \r\nindependently folding protein motif with 34-40 amino acid residues, as well as its function as a protein \r\ninteraction module with a flexible binding site, the WW domain was selected as a scaffold for the \r\ndesign of miniaturized proteins in this work.[10]\r\n\r\nTo establish the WW domain as a scaffold for miniprotein design, an iterative sequence analysis \r\napproach was initially employed to identify sequence-structure-stability-relationships, starting from a \r\nconsensus sequence of 90 native WW domains. In accordance with the identified sequence-structurestability relationships, three highly thermostable WW domain core scaffolds were designed, displaying \r\nmelting temperatures of 89 and 93 °C in thermal CD denaturation experiments. Multi-dimensional \r\nhigh-resolution NMR experiments of one highly thermostable scaffold peptide demonstrated the \r\ntypical WW domain structure, comprising three anti-parallel β-sheets and additional structural \r\nfeatures that contribute to its high thermostability. As a proof of concept for the design of functional \r\nminiproteins, binding motifs of the three different WW domain groups were grafted onto the \r\nthermostable WW domain core scaffold to obtain thermostable WW domains with group-specific \r\nbinding properties. Binding studies on the designed group-specific thermostable WW domains \r\nrevealed micromolar-to-sub-micromolar binding affinities to their respective ligands. In addition to the \r\nintroduction of native functions into the thermostable WW domain core scaffold, binding motifs for \r\nnon-native functions, such as metal- and organophosphate binding, were grafted onto a scaffold \r\npeptide. This resulted in the design of de novo thermostable WW domains with micromolar-tosubmicromolar binding affinities to their respective ligands. Following the successful design of an \r\nadenosine-triphosphate (ATP)-binding thermostable variant (WW-HS-ATP), further peptide variants \r\nwere designed with diverse sequence modifications to study the influence on binding affinity to ATP, \r\nADP and AMP. All de novo designed variants, as well as WW-HS-ATP, demonstrated micromolar binding \r\naffinities to ATP, ADP and AMP. To ascertain whether one of the ATP-binding peptides exhibits \r\nenzymatic activity in terms of phosphatase activity, exploratory studies were conducted, which \r\nidentified certain candidates that showed minimal phosphatase-like activity. To obtain thermostable \r\nWW domain miniproteins with enzymatic activity, further sequence redesign and additional studies \r\nare required. In conclusion, this work successfully demonstrated that the establishment of sequence-structure-stability relationships of native WW domains, resulted in the design of highly thermostable \r\nWW domain core scaffolds, which could be further functionalized with native and non-native \r\nfunctions, while maintaining thermostable properties. The designed thermostable WW domain \r\nscaffolds could serve as a template for future β-sheet miniprotein design with diverse receptor, sensor \r\nor catalytic properties."^^ . "2024" . . . . . . . "Christina Gabriele"^^ . "Lindner"^^ . "Christina Gabriele Lindner"^^ . . . . . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases (PDF)"^^ . . . "Diss_ChristinaLindner_21.08.24_final.pdf"^^ . . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases (Other)"^^ . . . . . . "indexcodes.txt"^^ . . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases (Other)"^^ . . . . . . "preview.jpg"^^ . . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases (Other)"^^ . . . . . . "medium.jpg"^^ . . . "The WW domain scaffold as model system for de novo design \r\nof miniaturized phosphate receptors and phosphatases (Other)"^^ . . . . . . "small.jpg"^^ . . "HTML Summary of #35610 \n\nThe WW domain scaffold as model system for de novo design \nof miniaturized phosphate receptors and phosphatases\n\n" . "text/html" . .