Simultaneously monitoring endogenous MAPK members in single living cells by multi-channel fluorescence correlation spectroscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Liyun Deng ◽  
Xiangyi Huang ◽  
Chaoqing Dong ◽  
Jicun Ren
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Mapk Pathway ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Mitogen Activated Protein Kinase ◽  
Fluorescence Correlation ◽  
Mitogen Activated Protein ◽  
Cellular Signal ◽  
Single Living Cells ◽  
Single Living

The mitogen-activated protein kinase (MAPK) pathway is a major module for cellular signal transduction. The dysregulation of MAPK pathway has been involved in the pathogenesis of multiple diseases ranging from...

scholarly journals Insulin-triggered repositioning of munc18c on syntaxin-4 in GLUT4 signalling

2008 ◽  
Vol 410 (2) ◽  
pp. 255-260 ◽  
Author(s):  
Natalie P. Smithers ◽  
Conrad P. Hodgkinson ◽  
Matt Cuttle ◽  
Graham J. Sale
Keyword(s):  
Plasma Membrane ◽  
Glucose Transporter ◽  
Cell Signalling ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Vesicle Docking ◽  
Fluorescence Correlation ◽  
Syntaxin 4 ◽  
Single Living Cells ◽  
Single Living

One of the most important actions of insulin is the stimulation of the uptake of glucose into fat and muscle cells. Crucial to this response is the translocation of GLUT4 (glucose transporter-4) to the plasma membrane. The insulin-stimulated GLUT4 vesicle docking at the plasma membrane requires an interaction between VAMP-2 (vesicle-associated membrane protein-2) on the GLUT4 vesicle and syntaxin-4 in the plasma membrane. In the basal state, munc18c is thought to preclude GLUT4 vesicle docking by inhibiting this interaction. Here, we have used FCS (fluorescence correlation spectroscopy) in single living cells to show that munc18c binds to syntaxin-4 in both the basal and insulin-stimulated states. We show that munc18c contains two binding sites for syntaxin-4, one of which is disrupted by insulin, while the other is activated by insulin. Insulin-triggered repositioning of munc18c on syntaxin-4 in this way in turn allows syntaxin-4 to adopt its ‘open’ conformation and bind VAMP-2, resulting in the docking of the GLUT4 vesicle at the cell surface. The results also demonstrate the utility of using FCS in intact single living cells to elucidate cell signalling events.

Mobility and distribution of replication protein A in living cells using fluorescence correlation spectroscopy

2007 ◽  
Vol 82 (2) ◽  
pp. 156-162 ◽  
Author(s):  
Christophe Braet ◽  
Holger Stephan ◽  
Ian M. Dobbie ◽  
Denisio M. Togashi ◽  
Alan G. Ryder ◽  
...  
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Protein A ◽  
Replication Protein A ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Replication Protein ◽  
Fluorescence Correlation
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