| Online-Ressource |
Verfasst von: | Neitz, Hermann [VerfasserIn]  |
| Paul, Niels Benjamin [VerfasserIn]  |
| Häge, Florian [VerfasserIn]  |
| Lindner, Christina [VerfasserIn]  |
| Graebner, Roman [VerfasserIn]  |
| Kovermann, Michael [VerfasserIn]  |
| Thomas, Franziska [VerfasserIn]  |
Titel: | Identification of novel functional mini-receptors by combinatorial screening of split-WW domains |
Verf.angabe: | Hermann Neitz, Niels Benjamin Paul, Florian R. Häge, Christina Lindner, Roman Graebner, Michael Kovermann and Franziska Thomas |
E-Jahr: | 2022 |
Jahr: | 14 Jul 2022 |
Umfang: | 12 S. |
Fussnoten: | Gesehen am 13.02.2023 |
Titel Quelle: | Enthalten in: Chemical science |
Ort Quelle: | Cambridge : RSC, 2010 |
Jahr Quelle: | 2022 |
Band/Heft Quelle: | 13(2022), 31, Seite 9079-9090 |
ISSN Quelle: | 2041-6539 |
Abstract: | β-Sheet motifs such as the WW domain are increasingly being explored as building blocks for synthetic biological applications. Since the sequence-structure relationships of β-sheet motifs are generally complex compared to the well-studied α-helical coiled coil (CC), other approaches such as combinatorial screening should be included to vary the function of the peptide. In this study, we present a combinatorial approach to identify novel functional mini-proteins based on the WW-domain scaffold, which takes advantage of the successful reconstitution of the fragmented WW domain of hPin1 (hPin1WW) by CC association. Fragmentation of hPin1WW was performed in both loop 1 (CC-hPin1WW-L1) and loop 2 (CC-hPin1WW-L2), and the respective fragments were linked to the strands of an antiparallel heterodimeric CC. Structural analysis by CD and NMR spectroscopy revealed structural reconstitution of the WW-domain scaffold only in CC-hPin1WW-L1, but not in CC-hPin1WW-L2. Furthermore, by using 1H-15N HSQC NMR, fluorescence and CD spectroscopy, we demonstrated that binding properties of fragmented hPin1WW in CC-hPin1WW-L1 were fully restored by CC association. To demonstrate the power of this approach as a combinatorial screening platform, we synthesized a four-by-six library of N- and C-terminal hPin1WW-CC peptide fragments that was screened for a WW domain that preferentially binds to ATP over cAMP, phophocholine, or IP6. Using this screening platform, we identified one WW domain, which specifically binds ATP, and a phosphorylcholine-specific WW-based mini-receptor, both having binding dissociation constants in the lower micromolar range. |
DOI: | doi:10.1039/D2SC01078J |
URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
kostenfrei: Volltext: https://doi.org/10.1039/D2SC01078J |
| kostenfrei: Volltext: https://pubs.rsc.org/en/content/articlelanding/2022/sc/d2sc01078j |
| DOI: https://doi.org/10.1039/D2SC01078J |
Datenträger: | Online-Ressource |
Sprache: | eng |
Bibliogr. Hinweis: | Forschungsdaten: Thomas, Franziska, 1980 - : Identification of novel functional mini-receptors by combinatorial screening of split-WW domains [research data] |
K10plus-PPN: | 1834593573 |
Verknüpfungen: | → Zeitschrift |
Identification of novel functional mini-receptors by combinatorial screening of split-WW domains / Neitz, Hermann [VerfasserIn]; 14 Jul 2022 (Online-Ressource)