I'm just wondering if anyone has any experience about the non-specific binding of GFP to the cytoskeleton, particularly actin fibers? Is it possible for the GFP protein to interact with actin cytoskeleton, not fusion protein but protein-protein interactions?
Thank you very much
Hi,
If you wonder because you have observed something look like co-localisation under fluorescent microscope, you should check the possibility that it could be due to "fluorescence breed-through (or "cross-talk)".
I would recommend to talk to a microscopy expert in your institute, if you have not (I believe you have one in the Beatson).
Nevertheless, what you see could be true.
I found this paper.
http://www.jbc.org/content/282/14/10465.full.pdf
What they found is, GFP interacts with myosin and inhibits myosin from interacting with actin. That's interesting, although the result does not necessarily predict whether you can expect GFP can be found at/near actin fibres.
Hi Shin-Ichiro,
I don't think its the breed through because the cell only express construct with GFP so there will only be GFP-fusion protein inside the cell and no other fluorescent molecule. In many of my images, it seems like GFP can somehow interact and non specifically bind to actin (it glows along the fiber (im not even sure if thats actin fiber but they look like so)). I dont think its the fused protein that makes this interaction, but i cannot 100% rule out this possibility.
Typically, GFP does not associate with the actin cytoskeleton. Yet, I've seen publications where GFP control showed partial "co-localization " with actin stress fibers. Since actin in these studies was stained with phalloidin, I assumed that what appeared to be a GFP signal, was bleeding from the phalloidin channel (see Dr. Higara's explanation above). It appears to be not your case. Since you have a fusion protein, it is rather likely that the non-GFP-part of it binds to actin. This is not surprising given that literally hundreds of proteins can bind to actin.
Dimitri Kudryshov: We are quite sceptical about this. When we look at the just GFP transfected cells, we kinda also see the colocalisation of GFP with actin however, so thats why we are not so sure. Hence I decided to ask if there's any known case that GFP can actually interact with actin either specifically or non specifically. Or maybe actin itself can self-fluoresce?
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