The intrinsically disordered TC‐1 interacts with Chibby via regions with high helical propensity

Gall, C. M.; Xu, Hanyu; Brickenden, Anne; Ai, Xuan; Choy, Wing Yiu

doi:10.1110/ps.073062707

Cited by 26 publications

(42 citation statements)

References 39 publications

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“…ProTα was purified using the method described by Yi et al [39]. The N-terminally His tagged TC-1 protein was extracted from inclusion bodies using 6 M guanidine hydrochloride and purified by affinity chromatography using Ni Sepharose™ 6 Fast Flow beads (Amersham Biosciences) [46]. The plasmid carrying the α-synuclein cDNA was kindly supplied by Dr. Pielak at the University of North Carolina-Chapel Hill.…”

Section: Methodsmentioning

confidence: 99%

“…It competes with β-catenin on binding to Chibby (Cby) and therefore inhibits the antagonistic action of Cby on β-catenin mediated transcription [44], [45]. Even though TC-1 is classified as an IDP, structural characterization shows that while the N-terminal half of the protein is largely unstructured, high helical propensity is present in the C-terminal part [42], [46]. α-synuclein, a well-studied IDP that has been found to be the main structural component of Lewy body fibrils found in patients with Parkinson’s disease [47], was also included in this study to add additional depth to our approach.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Molecular Crowding on the Dynamics of Intrinsically Disordered Proteins

2012

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Inside cells, the concentration of macromolecules can reach up to 400 g/L. In such crowded environments, proteins are expected to behave differently than in vitro. It has been shown that the stability and the folding rate of a globular protein can be altered by the excluded volume effect produced by a high density of macromolecules. However, macromolecular crowding effects on intrinsically disordered proteins (IDPs) are less explored. These proteins can be extremely dynamic and potentially sample a wide ensemble of conformations under non-denaturing conditions. The dynamic properties of IDPs are intimately related to the timescale of conformational exchange within the ensemble, which govern target recognition and how these proteins function. In this work, we investigated the macromolecular crowding effects on the dynamics of several IDPs by measuring the NMR spin relaxation parameters of three disordered proteins (ProTα, TC1, and α-synuclein) with different extents of residual structures. To aid the interpretation of experimental results, we also performed an MD simulation of ProTα. Based on the MD analysis, a simple model to correlate the observed changes in relaxation rates to the alteration in protein motions under crowding conditions was proposed. Our results show that 1) IDPs remain at least partially disordered despite the presence of high concentration of other macromolecules, 2) the crowded environment has differential effects on the conformational propensity of distinct regions of an IDP, which may lead to selective stabilization of certain target-binding motifs, and 3) the segmental motions of IDPs on the nanosecond timescale are retained under crowded conditions. These findings strongly suggest that IDPs function as dynamic structural ensembles in cellular environments.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effects of Molecular Crowding on the Dynamics of Intrinsically Disordered Proteins

2012

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“…It is a small protein of 106 amino acids without known functional domain but a nuclear localization signal and a leucin zipper-like sequence (10). It is rapidly degraded through ubiquitin-proteasome pathway (10), and is suggested to be a natively disordered protein (6,11).…”

mentioning

confidence: 99%

TC1(C8orf4) Is a Novel Endothelial Inflammatory Regulator Enhancing NF-κB Activity

Kim¹,

Kim²,

Kim

et al. 2009

The Journal of Immunology

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Endothelial inflammation is regulated by a complex molecular mechanism. TC1(C8orf4) is a novel regulator implicated in cancer and inflammation. It is a small protein conserved well among vertebrates. In zebrafish embryos, it is mostly expressed in angio-hematopoietic system and the overexpression induces edema. In human aortic endothelial cells and umbilical vein endothelial cells, TC1 transfection up-regulates key inflammatory cytokines, enzymes, and adhesion proteins including IL-6, IL-1α, COX-2, CXCL1, CCL5, CCL2, IL-8, ICAM1, VCAM1, and E-selectin, while TC1 knockdown down-regulates them. TC1 also enhances inflammatory parameters such as monocyte-endothelial adhesion and endothelial monolayer permeability. TC1 is up-regulated by IL-1β, TNF-α, LPS, and phorbol ester, and the up-regulation is inhibited by I-κB-kinase inhibitors. TC1, in turn, enhances the nuclear translocation of RelA and the DNA binding activity, suggesting a biological role of amplifying NF-κB signaling via a positive feedback. Our findings suggest that TC1 is a novel endothelial inflammatory regulator that might be implicated in inflammatory vascular diseases.

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“…In a previous study, we have shown that disordered TC-1 binds to Cby via its highly helical C-terminal region (18). Here we have carried out an NMR binding experiment by which unlabeled TC-1 was added to the 15 N-labeled Cby.…”

Section: Resultsmentioning

confidence: 99%

Structural Characterization of Partially Disordered Human Chibby: Insights into Its Function in the Wnt-Signaling Pathway

Mokhtarzada

Brickenden

et al. 2011

Biochemistry

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The Wnt/β-catenin signaling pathway is critical to embryonic development as well as adult tissue regeneration. Dysregulation of this pathway can lead to a variety of human diseases, in particular cancers. Chibby (Cby), a small and highly conserved protein, plays an antagonistic role in Wnt signaling by inhibiting the binding of β-catenin to Tcf/Lef family proteins, a protein interaction that is essential for the transcriptional activation of Wnt target genes. Cby is also involved in regulating intracellular distribution of β-catenin. Phosphorylated Cby forms a ternary complex with 14-3-3 protein and β-catenin, facilitating the export of β-catenin from the nucleus. On the other hand, the antagonistic function of Cby is inhibited upon binding to thyroid cancer-1 (TC-1). To dissect the structure−function relationship of Cby, we have used NMR spectroscopy, ESI-MS, CD, and DLS to extensively characterize the structure of human Cby. Our results show that the 126-residue Cby is partially disordered under nondenaturing conditions. While the N-terminal portion of the protein is predominantly unstructured in solution, the C-terminal half of Cby adopts a coiled-coil structure through self-association. Initial data for the binding studies of Cby to 14-3-3ζ (one of the isoforms in the 14-3-3 family) and TC-1 via these two distinct structural modules have also been obtained. It is noteworthy that in a recent large-scale analysis of the intrinsically disordered proteome of mouse, a substantial number of disordered proteins are predicted to have coiled-coil motif presence in their sequences. The combination of these two molecular recognition features could facilitate disordered Cby in assembling protein complexes via different modes of interaction.

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The intrinsically disordered TC‐1 interacts with Chibby via regions with high helical propensity

Cited by 26 publications

References 39 publications

Effects of Molecular Crowding on the Dynamics of Intrinsically Disordered Proteins

Effects of Molecular Crowding on the Dynamics of Intrinsically Disordered Proteins

TC1(C8orf4) Is a Novel Endothelial Inflammatory Regulator Enhancing NF-κB Activity

Structural Characterization of Partially Disordered Human Chibby: Insights into Its Function in the Wnt-Signaling Pathway

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