Source 1primary paper2004Science
Toolkit/reversible protein highlighting
reversible protein highlighting
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Reversible protein highlighting is a light-based live-cell imaging assay that uses a photochromic fluorescent protein to repeatedly highlight, erase, and re-highlight labeled molecules without destructive readout. It was applied to visualize stimulus-dependent, bidirectional nucleocytoplasmic shuttling of extracellular signal-regulated kinase (ERK) across the nuclear envelope.
Usefulness & Problems
Why this is useful
This method is useful for directly observing regulated protein movement in living cells when repeated marking of the same labeled population is required. The cited study specifically used it to monitor fast signaling-associated nucleocytoplasmic transport of ERK.
Source:
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Problem solved
It addresses the problem of visualizing rapid, regulated, bidirectional trafficking of signaling proteins across the nuclear envelope in a nondestructive manner. The evidence specifically supports its use for stimulus-dependent ERK shuttling.
Problem links
Need conditional control of signaling activity
DerivedReversible protein highlighting is a light-based functional imaging method that uses a photochromic fluorescent protein to repeatedly highlight, erase, and re-highlight labeled molecules in a nondestructive manner. It was used to directly visualize stimulus-dependent, bidirectional nucleocytoplasmic shuttling of extracellular signal-regulated kinase (ERK) across the nuclear envelope.
Need precise spatiotemporal control with light input
DerivedReversible protein highlighting is a light-based functional imaging method that uses a photochromic fluorescent protein to repeatedly highlight, erase, and re-highlight labeled molecules in a nondestructive manner. It was used to directly visualize stimulus-dependent, bidirectional nucleocytoplasmic shuttling of extracellular signal-regulated kinase (ERK) across the nuclear envelope.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
photochromic optical switchingphotochromic optical switchingreversible fluorescence highlightingreversible fluorescence highlightingTechniques
Functional AssayTarget processes
signalingInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: sensor
Implementation requires a photochromic fluorescent protein and light-based live-cell fluorescence imaging. The available evidence does not specify excitation wavelengths, protein fusion design, expression system, or instrumentation parameters.
The supplied evidence is limited to a single cited study and a single demonstrated application involving ERK nucleocytoplasmic shuttling. No quantitative performance metrics, spectral details, construct architecture, or broader validation across proteins, cell types, or organisms are provided here.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug reversible-protein-highlighting
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Source:
method capabilitysupports
Because of its photochromic properties, the fluorescent protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.
Source:
visualization capabilitysupports
Reversible protein highlighting was used to visualize stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting.
Source:
Comparisons
Source-backed strengths
The reported strength is reversible, nondestructive optical highlighting enabled by photochromic fluorescence, allowing repeated highlight-erase-rehighlight cycles. In the cited application, this enabled direct visualization of stimulus-dependent acceleration of bidirectional ERK flow across the nuclear envelope.
Compared with cDNA microarray
reversible protein highlighting and cDNA microarray address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling; same primary input modality: light
Compared with IRAP-pHluorin translocation assay
reversible protein highlighting and IRAP-pHluorin translocation assay address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling; same primary input modality: light
reversible protein highlighting and light-induced Fourier transform infrared (FTIR) difference spectroscopy address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling; same primary input modality: light
Ranked Citations
- 1.