Difference between revisions of "Part:BBa K3009020"

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The presence of FPR2 in transfected cells can be checked by flurescence microscopy and FACS. If no anti FPR2 antibody is available a high resolution microscope can even help to determine if FPR2 is located at the membrane, relying on the GFP reporter.
 
The presence of FPR2 in transfected cells can be checked by flurescence microscopy and FACS. If no anti FPR2 antibody is available a high resolution microscope can even help to determine if FPR2 is located at the membrane, relying on the GFP reporter.
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[[File:T--Freiburg--FPR2 calcium.png]]
  
 
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<figcaption> <b>Fig.2:</b> Calcium release of transfected FPR2-eGFP HEK cells. A) PSMa3 toxin and activator WKYMVm (B) leads to calcium flux
 
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Revision as of 20:49, 20 October 2019


FPR2-receptor with eGFP

The human formyl peptide receptor 2 (FPR2) is a G-protein coupeled receptor which is physiologically expressed on immune cell lineages like neutrophils and T-cells. Amongst other peptides the FPR2 senses the staphylococcus aureus toxin PSMa3. It was suggested by Cheung et al 2014 that the binding mechanism relies on the formylated N-terminus of the peptide as well as on the C-terminus. In response to receptor activation FPR2 elicits a signalling cascade depending on calcium ions as second messengers. Ultimately this leads to immune cell activation, secretion of inflammatory cytokines and chemotaxis. All of this is connected with an inflammatory outcome in vivo. The neutrophil activation by FPR2 is therefore an important mechanism of its toxicity because it leads to an aggravation of the inflammation related symptoms in Staphylococcus aureus infection

This Biobrick can be used in signalling studies or the cellular detection of several small formylated amyloidogenic peptides. As a controls two peptides with inhibitry (WRWW4) and activating (WKYMVm) effect on FPR2 are availabe.

The presence of FPR2 in transfected cells can be checked by flurescence microscopy and FACS. If no anti FPR2 antibody is available a high resolution microscope can even help to determine if FPR2 is located at the membrane, relying on the GFP reporter.


File:T--Freiburg--FPR2 calcium.png

Fig.2: Calcium release of transfected FPR2-eGFP HEK cells. A) PSMa3 toxin and activator WKYMVm (B) leads to calcium flux

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1063
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]