Dissection of Kinase Isoforms through Orthogonal and Chemical Inducible Signaling Cascades

Zheng, Siqi; Fan, Xinyuan; Wang, Jie; Zhao, Jingyi; Chen, Peng R.

doi:10.1002/cbic.201700255

Cited by 10 publications

(6 citation statements)

References 31 publications

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“…We then employed a luciferase reporter, SRE-luc, to monitor OspF activity in living cells. 31 SRE-luc reporter responds to ERK/MAPK signaling, allowing the luciferase expression level correlated with the endogenous ERK activity. For example, the reactivated OspF* attenuated ERK activity constitutively and thus reduced luciferase expression and the bioluminescence signal.…”

Section: Resultsmentioning

confidence: 99%

“…First, we showed that, upon photoactivation of OspF* inside cells, the attenuation of ERK/p38 phosphorylation was observed as fast as 5 min (Figure S8) while the protein level of OspF* was not changed even 60 min after photoactivation. We then employed a luciferase reporter, SRE-luc, to monitor OspF activity in living cells . SRE-luc reporter responds to ERK/MAPK signaling, allowing the luciferase expression level correlated with the endogenous ERK activity.…”

Section: Resultsmentioning

confidence: 99%

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Bioorthogonal Engineering of Bacterial Effectors for Spatial–Temporal Modulation of Cell Signaling

et al. 2018

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The complicated and entangled cell signaling network is dynamically regulated by a wide array of enzymes such as kinases. It remains desirable but challenging to specifically modulate individual, endogenous kinases within a cell, particularly in a spatial–temporally controlled fashion. Current strategies toward regulating the intracellular functions of a kinase of interest either lack specificity or require genetic engineering that may perturb its physiological activity. Herein, we harnessed a bacterial effector OspF for optical and chemical modulation of the endogenous mitogen-activated protein kinase (MAPK) cascade in living cells and mice. The phospho-lyase OspF provided high specificity and spatial resolution toward the desired kinase such as the extracellular signal-regulated kinase (ERK), while the genetically encoded bioorthogonal decaging strategy enabled its temporal activation in living systems. The photocaged OspF (OspF*) was applied to dissect the subcellular signaling roles of ERK in nucleus as opposed to cytoplasm, while the chemically caged OspF (OspFc) was introduced into living mice to modulate ERK-mediated gene expression. Finally, our spatially and chemically controlled OspFc was further used to precisely tune immune responses in T cells. Together, our bioorthogonal engineering strategy on bacterial effectors offers a general tool to modulate cell signaling with high specificity and spatial–temporal resolution.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Bioorthogonal Engineering of Bacterial Effectors for Spatial–Temporal Modulation of Cell Signaling

et al. 2018

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“…By using trans ‐cyclooctene‐caged‐lysine groups (Scheme B), it was possible not only to control the function of proteins in cells but also in live mice . This chemistry also enabled kinase signaling pathways in cells to be controlled …”

Section: Applications Of Dissociative Bioorthogonal Reactions In the mentioning

confidence: 99%

“…[84] This chemistry also enabledk inase signaling pathways in cells to be controlled. [85] It is also possible to control cell-cell communication by bioorthogonal release reactions. An epitope with ac aged azidolysine residue was loaded onto major histocompatibility complex class I( MHC-I) on dendritic cells.…”

Section: Using Dissociative Chemistry To Control the Activityo F Protmentioning

confidence: 99%

Dissociative Bioorthogonal Reactions

Franzini

2019

ChemBioChem

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Bioorthogonal reactions that proceed readily under physiological conditions without interference from biomolecules have found widespread application in the life sciences. Complementary to the bioorthogonal reactions that ligate two molecules, reactions that release a molecule or cleave a linker are increasingly attracting interest. Such dissociative bioorthogonal reactions have a broad spectrum of uses, for example, in controlling bio‐macromolecule activity, in drug delivery, and in diagnostic assays. This review article summarizes the developed bioorthogonal reactions linked to a release step, outlines representative areas of the applications of such reactions, and discusses aspects that require further improvement.

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“…For example, several protecting groups can be removed bioorthogonally inside live mammalian cells, and these chemistries have been used to switch on protein function by genetic code expansion. Intracellular bioorthogonal reactions that have been used in this purpose include inverse electron demand Diels–Alder reactions [18,59–61], 1,3-dipolar cycloadditions [62], Staudinger reactions [63], and palladium-catalysed propargyl removal (Table 4) [57]. Currently, all of these have only been demonstrated in caged lysine molecules ( 61 , 70 , 71 , 85 ) through a number of examples, including activation of luciferases, kinases, nucleases etc.…”

Section: Small-molecule Induced Activation or Inhibitionmentioning

confidence: 99%

Using genetically incorporated unnatural amino acids to control protein functions in mammalian cells

Nödling

Spear

Williams

et al. 2019

Essays in Biochemistry

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Genetic code expansion allows unnatural (non-canonical) amino acid incorporation into proteins of interest by repurposing the cellular translation machinery. The development of this technique has enabled site-specific incorporation of many structurally and chemically diverse amino acids, facilitating a plethora of applications, including protein imaging, engineering, mechanistic and structural investigations, and functional regulation. Particularly, genetic code expansion provides great tools to study mammalian proteins, of which dysregulations often have important implications in health. In recent years, a series of methods has been developed to modulate protein function through genetically incorporated unnatural amino acids. In this review, we will first discuss the basic concept of genetic code expansion and give an up-to-date list of amino acids that can be incorporated into proteins in mammalian cells. We then focus on the use of unnatural amino acids to activate, inhibit, or reversibly modulate protein function by translational, optical or chemical control. The features of each approach will also be highlighted.

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Dissection of Kinase Isoforms through Orthogonal and Chemical Inducible Signaling Cascades

Cited by 10 publications

References 31 publications

Bioorthogonal Engineering of Bacterial Effectors for Spatial–Temporal Modulation of Cell Signaling

Bioorthogonal Engineering of Bacterial Effectors for Spatial–Temporal Modulation of Cell Signaling

Dissociative Bioorthogonal Reactions

Using genetically incorporated unnatural amino acids to control protein functions in mammalian cells

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