Phosphorylation Reduces the Mechanical Stability of the α‐Catenin/ β‐Catenin Complex

Le, Shimin; Yu, Miao; Yan, Jie

doi:10.1002/ange.201911383

Cited by 5 publications

(6 citation statements)

References 24 publications

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“…Mechanotransduction is based on the binding and unbinding of factors that are influenced by force and occur within a time frame ranging from seconds to minutes depending on various factors such as the concentration of binding factors and the strength of interactions (Supporting Information S12). − The significance of a few seconds lifetime over pN forces lies in two reasons: First, pN forces are necessary to activate mechanosensing domains in various mechanosensing proteins, such as talin rod domains, α-catenins, , and FLNC rod-2 domains shown in this study, which requires seconds to reach physiologically relevant loading rates on the order of pN per second. Second, the binding of signaling proteins to activated mechanosensing proteins takes time.…”

Section: Discussionmentioning

confidence: 94%

Force-Dependent Structural Changes of Filamin C Rod Domains Regulated by Filamin C Dimer

Deng

Yan

2023

J. Am. Chem. Soc.

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Filamin C (FLNC), a large dimeric actin-binding protein in muscle cells, plays a critical role in transmitting force in the cytoskeleton and that between membrane receptors and the cytoskeleton. It performs crucial mechanosensing and downstream mechanotransduction functions via force-dependent interactions with signaling proteins. Mutations in FLNC have been linked to muscle and heart diseases. The mechanical responses of the force-bearing elements in FLNC have not been determined. This study investigated the mechanical responses of FLNC domains and their dimerization interface using magnetic tweezers. Results showed high stability of the N-terminal domains in the rod-1 segment but significant changes in the rod-2 domains in response to forces of a few piconewtons (pN). The dimerization interface, formed by the R24 domain, has a lifetime of seconds to tens of seconds at pN forces, and it dissociates within 1 s at forces greater than 14 pN. The findings suggest the FLNC dimerization interface provides sufficient mechanical stability that enables force-dependent structural changes in rod-2 domains for signaling protein binding and maintains structural integrity of the rod-1 domains.

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Section: Discussionmentioning

confidence: 94%

Force-Dependent Structural Changes of Filamin C Rod Domains Regulated by Filamin C Dimer

Deng

Yan

2023

J. Am. Chem. Soc.

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“…A vertical magnetic tweezers setup was combined with a disturbance-free, rapid solution-exchange flow channel for conducting in vitro protein stretching experiments [22][23][24]. All in vitro protein stretching experiments were performed in solution containing: 1X PBS, 1% BSA, 2 mM DTT, 10 mM sodium L-ascorbate at 22 ± 1 o C. The force calibration of the magnetic tweezers setup has a 10% uncertainty due to the heterogeneity of the diameter of paramagnetic beads [22] and the bead height determination of the magnetic tweezers setup has a ⇠ 2-5 nm uncertainty due to the thermal fluctuation of the tethered bead and the resolution of the camera [10]. ); N =67, F = 16.3±1.7 pN, H=117.3±10.9 nm (5.0 pN s -1 ); (i).…”

Section: Methodsmentioning

confidence: 99%

“…We utilized a single-molecule construct that converts a protein-protein interaction into an intra-molecular interaction (Figure 4a) [10,11,26,[32][33][34]. Briefly, the construct of the HMP1-HMP2 interface consists of the N12 domain of HMP1, the Nt peptide of HMP2, and a long flexible unstructured peptide linker in between.…”

Section: Hmp1-hmp2 Interface Provides Su Cient Mechanical Stability For Supporting Tension Transmissionmentioning

confidence: 99%

“…The tension in the linkage has been estimated to be in the order of several piconewtons (pN) by molecular tension sensors [2][3][4]. Signalling proteins interact with mechanosensing proteins in the linkage in a force-dependent manner (e.g., vinculin-↵-catenin interaction), regulating the strength of the adherens junction and converting the mechanical cues into a cascade of downstream biochemical signalling events [5][6][7][8][9][10][11][12]. While the mechano-biochemical regulations of the mammalian adherens junction have been intensively investigated over recent years [4,[6][7][8][9][10][11][12], its counterpart in C. elegans, an important model system for development and aging, has been much less understood [2][3][4].…”

Section: Introductionmentioning

confidence: 99%

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Mechano-Biochemical Regulation of the C. elegans HMP1–HMP2 protein complex

Martin

et al. 2021

Preprint

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The HMP1–HMP2 protein complex, a counterpart of α-catenin–β-catenin complex in C. elegans, mediates the tension transmission between HMR1 (cadherin) and actin cytoskeleton and serves as a critical mechanosensor at the cell–cell adherens junction. The complex has been shown to play critical roles in embryonic development and tissue integrity in C. elegans. The complex is subject to tension due to internal actomyosin contractility and external mechanical micro-environmental perturbations. However, how tension regulates the stability and interaction of HMP1–HMP2 complex has yet to be investigated. Here, we directly quantify the mechanical stability of the full-length HMP1 and its force-bearing modulation domains (M1-M3), and show that they unfold within physiological level of tension (pico-newton scale). The inter-domain interactions within the modulation domain leads to strong mechanical stabilization of M1 in HMP1, resulting in a significantly stronger force threshold to expose the buried vinculin binding site compared to the M1 domain in α-catenins. Moreover, we also quantify the mechanical stability of the intermolecular HMP1–HMP2 interface and show that it is mechanically stable enough to support the tension-transmission and tension-sensing of the HMP1 modulation domains. Further, we show that single-residue phosphomimetic mutation (Y69E) on HMP2 weakens the mechanical stability of the HMP1–HMP2 interface and thus weakens the force-transmission molecular linkage and the associated mechanosensing functions. Together, these results provide a mechano-biochemical understanding of C. elegans HMP1–HMP2 protein complex's roles in mechanotransduction.

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“…Using its ultrasharp AFM tip, it is able not only to image nanoscale surface but also to manipulate a single molecule mechanically as single-molecule force spectroscopy (SMFS) [8][9][10][11][12]. By stretching one molecule under defined coordination, SMFS can induce the conformational change of a molecule such as protein (un)folding [13][14][15], breaking a stable chemical bond as well as capturing important transient states [16][17][18][19][20][21]. For example, several metal-binding proteins have been studied by AFM-SMFS, in which the metal cluster is ruptured during protein unfolding [22][23][24][25][26][27][28][29].…”

Section: Introductionmentioning

confidence: 99%

Highly Dynamic Polynuclear Metal Cluster Revealed in a Single Metallothionein Molecule

et al. 2021

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Human metallothionein (MT) is a small-size yet efficient metal-binding protein, playing an essential role in metal homeostasis and heavy metal detoxification. MT contains two domains, each forming a polynuclear metal cluster with an exquisite hexatomic ring structure. The apoprotein is intrinsically disordered, which may strongly influence the clusters and the metal-thiolate (M-S) bonds, leading to a highly dynamic structure. However, these features are challenging to identify due to the transient nature of these species. The individual signal from dynamic conformations with different states of the cluster and M-S bond will be averaged and blurred in classic ensemble measurement. To circumvent these problems, we combined a single-molecule approach and multiscale molecular simulations to investigate the rupture mechanism and chemical stability of the metal cluster by a single MT molecule, focusing on the Zn4S11 cluster in the α domain upon unfolding. Unusual multiple unfolding pathways and intermediates are observed for both domains, corresponding to different combinations of M-S bond rupture. None of the pathways is clearly preferred suggesting that unfolding proceeds from the distribution of protein conformational substates with similar M-S bond strengths. Simulations indicate that the metal cluster may rearrange, forming and breaking metal-thiolate bonds even when MT is folded independently of large protein backbone reconfiguration. Thus, a highly dynamic polynuclear metal cluster with multiple conformational states is revealed in MT, responsible for the binding promiscuity and diverse cellular functions of this metal-carrier protein.

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Phosphorylation Reduces the Mechanical Stability of the α‐Catenin/ β‐Catenin Complex

Cited by 5 publications

References 24 publications

Force-Dependent Structural Changes of Filamin C Rod Domains Regulated by Filamin C Dimer

Force-Dependent Structural Changes of Filamin C Rod Domains Regulated by Filamin C Dimer

Mechano-Biochemical Regulation of the C. elegans HMP1–HMP2 protein complex

Highly Dynamic Polynuclear Metal Cluster Revealed in a Single Metallothionein Molecule

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