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closeIs this action of GFP also true for other fluorescent proteins?
Posted by binxz on 09 Jan 2007 at 21:50 GMT
I am using the red fluorescent protein (RFP) as a reporter for eNOS in mesangial cells. Does the RFP affect the ubiquitination-proteosome pathways? Any comments to this questions would be greatly appreciated.
RE: Is this action of GFP also true for other fluorescent proteins?
mathijs replied to binxz on 10 Jan 2007 at 12:24 GMT
Don't know what you mean with RFP, is this DsRed? DsRed and the EGFP color variants EYFP-ECFP-EBFP all have the same effect on polyubiquitination.
RE: RE: Is this action of GFP also true for other fluorescent proteins?
Joshua replied to mathijs on 01 Feb 2007 at 23:41 GMT
To follow up Binxz's question:
Might many over-expressed proteins competitively inhibit and protein turnover? This paper holds out maxGFP as a negative control that does not inhibit ubiquitination. However, maxGFP is the only over-expressed protein in this paper that doesn’t have this effect. Maybe polyubiuitination inhibition is the general consequence of over-expression, and maxGFP is the exception! From the paper, it seems that beta-gal over-expression does not inhibit polyubiquitination, but what other reporters do?
RE: RE: RE: Is this action of GFP also true for other fluorescent proteins?
mathijs replied to Joshua on 14 Feb 2007 at 16:20 GMT
I don't think that maxGFP is the exception. If overexpression would affect polyubiquitination in general, averse effects would have been observed earlier. And overexpression of API2-MALT1 (and other molecules in the signaling cascade to NF-kB) increase polyubiquitination.