Diffusion of the disordered E-cadherin tail on β-catenin

Wiggers, Felix; Wohl, Samuel; Dubovetskyi, Artem; Rosenblum, Gabriel; Zheng, Wenwei; Hofmann, Hagen

doi:10.1101/2021.02.03.429507

Cited by 7 publications

(24 citation statements)

References 94 publications

(116 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This effect increases in magnitude with increasing ionic strength and is ablated when the five charged residues are mutated to alanine in the 5X construct. Intriguingly the results of the recent single-molecule study of the E-cadherin-βcatenin interaction led to a similar suggestion, whereby there is the continuous detachment and reattachment of local E-cadherin segments [57]. We share the authors' conclusion of a Velcro-like design of many weak contacts on top of a few persistent interactions.…”

Section: Discussionsupporting

confidence: 78%

“…Similar behaviour has been observed in the crystal structures of other β-catenin complexes, providing tantalising hints to the underlying mechanisms of recognition [10,21,23,[26][27][28]56]. Most recently, a single-molecule study by Hofmann and co-workers on the interaction between β-catenin and E-cadherin revealed a rugged energy landscape of E-cadherin with many shallow minima, a fuzzy complex, and a mechanism of intrachain diffusion of E-cadherin on β-catenin [57]. Here, we used site-directed mutagenesis and kinetic analysis to explore the significance of this plasticity in the β-catenin-TCF7L2 complex and to define the mechanism of association between the two proteins.…”

Section: Figuresupporting

confidence: 64%

“…Here, we used site-directed mutagenesis and kinetic analysis to explore the significance of this plasticity in the β-catenin-TCF7L2 complex and to define the mechanism of association between the two proteins. A-C) show the structures of TCF7L2 (red) in complex with β-catenin (cyan) obtained by Poy et al [22], Graham et al [57], and Sampietro et al [55], respectively. Structure (C) also has a peptide from the bound protein BCL9 (purple).…”

Section: Figurementioning

confidence: 95%

“…Schematics showing the crystal structures of the β-catenin-TCF7L2 complex. (A-C) show the structures of TCF7L2 (red) in complex with β-catenin (cyan) obtained by Poy et al [22], Graham et al[57], and Sampietro et al[55], respectively. Structure (C) also has a peptide from the bound protein BCL9 (purple).…”

mentioning

confidence: 98%

See 3 more Smart Citations

Parallel and Sequential Pathways of Molecular Recognition of a Tandem-Repeat Protein and Its Intrinsically Disordered Binding Partner

et al. 2021

View full text Add to dashboard Cite

The Wnt signalling pathway plays an important role in cell proliferation, differentiation, and fate decisions in embryonic development and the maintenance of adult tissues. The twelve armadillo (ARM) repeat-containing protein β-catenin acts as the signal transducer in this pathway. Here, we investigated the interaction between β-catenin and the intrinsically disordered transcription factor TCF7L2, comprising a very long nanomolar-affinity interface of approximately 4800 Å2 that spans ten of the twelve ARM repeats of β-catenin. First, a fluorescence reporter system for the interaction was engineered and used to determine the kinetic rate constants for the association and dissociation. The association kinetics of TCF7L2 and β-catenin were monophasic and rapid (7.3 ± 0.1 × 107 M−1·s−1), whereas dissociation was biphasic and slow (5.7 ± 0.4 × 10−4 s−1, 15.2 ± 2.8 × 10−4 s−1). This reporter system was then combined with site-directed mutagenesis to investigate the striking variability in the conformation adopted by TCF7L2 in the three different crystal structures of the TCF7L2–β-catenin complex. We found that the mutation had very little effect on the association kinetics, indicating that most interactions form after the rate-limiting barrier for association. Mutations of the N- and C-terminal subdomains of TCF7L2 that adopt relatively fixed conformations in the crystal structures had large effects on the dissociation kinetics, whereas the mutation of the labile sub-domain connecting them had negligible effect. These results point to a two-site avidity mechanism of binding with the linker region forming a “fuzzy” complex involving transient contacts that are not site-specific. Strikingly, the two mutations in the N-terminal subdomain that had the largest effects on the dissociation kinetics showed two additional phases, indicating partial flux through an alternative dissociation pathway that is inaccessible to the wild type. The results presented here provide insights into the kinetics of the molecular recognition of a long intrinsically disordered region with an elongated repeat-protein surface, a process found to involve parallel routes with sequential steps in each.

show abstract

Section: Discussionsupporting

confidence: 78%

Section: Figuresupporting

confidence: 64%

Section: Figurementioning

confidence: 95%

mentioning

confidence: 98%

See 2 more Smart Citations

Parallel and Sequential Pathways of Molecular Recognition of a Tandem-Repeat Protein and Its Intrinsically Disordered Binding Partner

et al. 2021

View full text Add to dashboard Cite

show abstract

“…[62] identified a fuzzy complex, where segments of E-cadherin (E-cad) diffuse dynamically royalsocietypublishing.org/journal/rsob Open Biol. 11: 210222 with high affinity over a large surface area of β-cat, in a manner distinct to the interactions observed between β-cat and Tcf4 [42][43][44]62]. β-cat is a cytoplasmic multifunctional repeat protein consisting of three domains: a central region domain, consisting of 12 imperfect armadillo domains which serve as the major interaction domain; a disordered N-terminal domain and a disordered CTD [62][63][64][65].…”

Section: Highly Dynamic E-cadherin Tail and β-Cat Interactionmentioning

confidence: 99%

Intrinsically disordered proteins: modes of binding with emphasis on disordered domains

2021

View full text Add to dashboard Cite

Our notions of protein function have long been determined by the protein structure–function paradigm. However, the idea that protein function is dictated by a prerequisite complementarity of shapes at the binding interface is becoming increasingly challenged. Interactions involving intrinsically disordered proteins (IDPs) have indicated a significant degree of disorder present in the bound state, ranging from static disorder to complete disorder, termed ‘random fuzziness’. This review assesses the anatomy of an IDP and relates how its intrinsic properties permit promiscuity and allow for the various modes of interaction. Furthermore, a mechanistic overview of the types of disordered domains is detailed, while also relating to a recent example and the kinetic and thermodynamic principles governing its formation.

show abstract