C-terminal fusion of eGFP to the bradykinin B-2 receptor strongly affects down-regulation but not receptor internalization or signaling

A functional comparison was made between the wildtype bradykinin B, receptor (B(2)wt) and the chimera B(2)eGFP (enhanced green-fluorescent protein fused to the C-terminus of B(2)Wt), both stably expressed in HEK 293 cells. There was almost no difference in terms of ligand-inducible receptor phosphorylation and internalization, signal transduction (accumulation of inositol phosphates) or expression and affinity. However, stimulation for up to 8 h with 10 mu M bradykinin (BK) resulted in a strong decrease in surface receptors (by 60% within 5 h) in B(2)Wt, but not in B(2)eGFP. When the expression levels of both constructs where comparably reduced using a weaker promoter, long-term stimulation resulted in a reduction in surface receptors for B(2)wt(low) to less than 20% within 1 h, whereas the chimera B(2)eGFP(low) still displayed 50% binding activity after 2 h. A 1-h incubation in the absence of BK resulted in a recovery of 60% of the binding in B(2)wt(low) after 1-h stimulation with BK, but of only 20% after 7-h stimulation. In contrast, B(2)eGFP(low) levels were restored to more than 70%, even after 7-h stimulation. These data indicate that although the fusion of eGFP to B(2)wt does not affect its ligand-induced internalization, it strongly reduces the down-regulation, most likely by promoting receptor recycling over degradation

The process gg -> WW as a background to the Higgs signal at the LHC

The production of W pairs from the one-loop gluon fusion process is studied. Formulas are presented for the helicity amplitudes keeping the top mass finite, but all other quark masses zero. The correlations among the leptons coming from the W bosons are kept. The contribution of this background to the Higgs boson search in the WW decay mode at the LHC is estimated by applying the cuts foreseen in experimental searches using the PYTHIA Monte Carlo program. Kinematic distributions for the final state leptons are compared to those of the Higgs boson signal and of the q qbar -> WW background. After applying final cuts, the gg background is found to be large, at the level of 35% of the q qbar background.The characteristics of the gg background are very similar to those of the signal. Therefore, an experimental normalization of this background component appears to be very difficult and the uncertainty must largely be determined by theory. As a result, the significance of a Higgs signal in the gg -> H -> WW mode at the LHC is reduced.Comment: 24 pages, 4 figure