Molecular Mechanism of Inward Rectifier Potassium Channel 2.3 Regulation by Tax-Interacting Protein-1

Yan, Xiao‐Jie; Zhou, Hao; Zhang, Jinxiu; Shi, Chaowei; Xie, Xiao‐Bi; Wu, Yafang; Tian, Changlin; Shen, Yuequan; Long, Jiafu

doi:10.1016/j.jmb.2009.07.060

Cited by 35 publications

(63 citation statements)

References 46 publications

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“…The protein used for hydrogel formation should be easily purified and have an impressively high yield, such as that of TIP1 [26]. After ensuring that no endogenous cysteine was present and analysing the protein’s crystal structure [26], we genetically modified four amino acids to cysteine at the outer surface (T10C, S42C, S101C and S113C) for further hydrogelation (Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…After ensuring that no endogenous cysteine was present and analysing the protein’s crystal structure [26], we genetically modified four amino acids to cysteine at the outer surface (T10C, S42C, S101C and S113C) for further hydrogelation (Figure 1). The TIP1 4C protein yield was comparable to that of TIP1; however, if the purified TIP1 4C (Figure S1) was placed at room temperature for approximately 15 minutes, it was unstable and it precipitated, especially when TIP1 4C was present at a high concentration (approximately 10 mg/ml).…”

Section: Resultsmentioning

confidence: 99%

“…Such cases are surely limiting the range of proteins that can be chosen for hydrogel formation and subsequent applications. In order to overcome that problem,we rationally replaced two or four amino acids with cysteine in TIP1 with no inherent cysteines [26] according to its crystal structure and used rheology and SEM to characterise the corresponding hydrogels that were formed by thiol-maleimide reaction. We also incorporated a bioactive peptide, GRGDSP, at the C-terminus of TIP1 2C and uncovered its roles in cell spreading and proliferation in the gel by Live-dead assay and CCK-8 assay.…”

Section: Introductionmentioning

confidence: 99%

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A Genetically Modified Protein-Based Hydrogel for 3D Culture of AD293 Cells

Wang

Diao

et al. 2014

PLoS ONE

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Hydrogels have strong application prospects for drug delivery, tissue engineering and cell therapy because of their excellent biocompatibility and abundant availability as scaffolds for drugs and cells. In this study, we created hybrid hydrogels based on a genetically modified tax interactive protein-1 (TIP1) by introducing two or four cysteine residues in the primary structure of TIP1. The introduced cysteine residues were crosslinked with a four-armed poly (ethylene glycol) having their arm ends capped with maleimide residues (4-armed-PEG-Mal) to form hydrogels. In one form of the genetically modification, we incorporated a peptide sequence ‘GRGDSP’ to introduce bioactivity to the protein, and the resultant hydrogel could provide an excellent environment for a three dimensional cell culture of AD293 cells. The AD293 cells continued to divide and displayed a polyhedron or spindle-shape during the 3-day culture period. Besides, AD293 cells could be easily separated from the cell-gel constructs for future large-scale culture after being cultured for 3 days and treating hydrogel with trypsinase. This work significantly expands the toolbox of recombinant proteins for hydrogel formation, and we believe that our hydrogel will be of considerable interest to those working in cell therapy and controlled drug delivery.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Genetically Modified Protein-Based Hydrogel for 3D Culture of AD293 Cells

Wang

Diao

et al. 2014

PLoS ONE

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“…5b) and further suggested that the phosphorylation of Thr-351 could have a role in modulating Tax-1 binding to its PDZ-containing partners. The importance of PBM phosphorylation is strengthened by recent evidence showing that the association of Taxinteracting protein-1 (TIP-1), through its PDZ domain, with Kir2.3 PBM was abolished by its phosphorylation inside the cell [34]. Nevertheless, it appears that the regulation of PDZ-based interactions by phosphorylation of C-terminal PBMs [35] is presenting a complex pattern, since the effect of this post-translational modification varies from a disruption of the interaction between PDZ domain and protein partners [34] to an increase in the binding ability [36].…”

Section: Discussionmentioning

confidence: 99%

“…The importance of PBM phosphorylation is strengthened by recent evidence showing that the association of Taxinteracting protein-1 (TIP-1), through its PDZ domain, with Kir2.3 PBM was abolished by its phosphorylation inside the cell [34]. Nevertheless, it appears that the regulation of PDZ-based interactions by phosphorylation of C-terminal PBMs [35] is presenting a complex pattern, since the effect of this post-translational modification varies from a disruption of the interaction between PDZ domain and protein partners [34] to an increase in the binding ability [36]. Notably, Ser/Thr substitutions with carboxylic side chains of Glu or Asp are not expected to display a phosphomimetic effect [37], although the steric hindrance as well as the charge of these modified residues could affect all the involved interactions, like those occurring between Tax-1 PBM and PDZ partners.…”

Section: Discussionmentioning

confidence: 99%

The pleiotropic protein kinase CK2 phosphorylates HTLV-1 Tax protein in vitro, targeting its PDZ-binding motif

et al. 2010

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The HTLV-1 transactivator Tax is an oncoprotein capable of deregulating the expression of many cellular genes and interfering with signalling pathways. Here we show that Tax-1 is phosphorylated in vitro by the pleiotropic human serine/threonine kinase CK2 at three residues, Ser-336, Ser-344 and Thr-351, close to and within its C-terminal PDZ-binding motif. We also show that the mutation of Thr-351 to aspartate abolishes Tax-1 binding to the scaffold protein hDlg, a tumour suppressor factor, while having no effect on transactivation. These results suggest that CK2, whose constitutive activity is often hijacked by viruses to sustain their vital cycle, could modulate Tax-1 oncogenic interactions.

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Rational Design of Multifunctional Hetero‐Hexameric Proteins for Hydrogel Formation and Controlled Delivery of Bioactive Molecules

Zhang

Zhou

Xie

et al. 2014

Adv Healthcare Materials

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A hetero-hexameric protein system is developed in this study, which not only functions as cross-linkers for hydrogel formation but also offers docking sites for controlled delivery of bioactive molecules. First, a hexameric protein with two, four, and six tax-interacting protein-1 (TIP-1), respectively (named as 2T, 4T, and 6T), is designed and obtained. As the hexapeptide ligand (WRESAI) can specifically bind to TIP-1 with high affinity, the hexameric proteins of 2T, 4T, and 6T can be used to crosslink the self-assembling nanofibers of Nap-GFFYGGGWRESAI, leading to formation of injectable biohybrid hydrogels with tunable mechanical properties. Furthermore, a hetero-hexameric protein containing four TIP-1 and two C-terminal moiety of the pneumococcal cell-wall amidase LytA (C-LytA) proteins is designed and engineered (named as 4T2C). The 4T2C proteins can not only serve as cross-linkers for hydrogel formation but also provide docking sites for loading and controlled release of model drug Rhoda-GGK'. This study opens up new opportunities for further development of multifunctional hetero- recombinant protein-based hydrogels for biological applications.

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Molecular Mechanism of Inward Rectifier Potassium Channel 2.3 Regulation by Tax-Interacting Protein-1

Cited by 35 publications

References 46 publications

A Genetically Modified Protein-Based Hydrogel for 3D Culture of AD293 Cells

A Genetically Modified Protein-Based Hydrogel for 3D Culture of AD293 Cells

The pleiotropic protein kinase CK2 phosphorylates HTLV-1 Tax protein in vitro, targeting its PDZ-binding motif

Rational Design of Multifunctional Hetero‐Hexameric Proteins for Hydrogel Formation and Controlled Delivery of Bioactive Molecules

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