Description
Receptor activity-modifying proteins (RAMPs) are a class of protein which interact with and modulate the activities of several Class B G Protein-Coupled Receptors including the receptors for secretin, calcitonin (CT), glucagon, and vasoactive intestinal peptide (VIP).There are three distinct types of RAMPs, designated RAMP1, RAMP2, and RAMP3, each encoded by a separate gene.
Currently the function of RAMPs is divided into 2 class activities. Association of RAMPs with either the CT or CALCR proteins forms 6 different receptors from the calcitonin receptor family. When associated with the Calcitonin receptor (CTR) or Calcitonin receptor-like (CALCRL) RAMPs can change the selectivity of the receptor for a specific hormone. In the cases of the other receptors mentioned however, there is no evidence that they can do this, but instead function to regulate trafficking of receptors from the ER / golgi to the membrane.
GenScript's cloned human AMY2 (RAMP2 + CALCR)–expressing cell line is generated in the CHO-K1/Gα15 host.
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| Culture Medium |
Ham’s F-12K (Kaighn’s), 10% FBS, 100 μg/ml Hygromycin B (Cat. #10687010, Invitrogen), 200 μg/ml Zeocin (Cat. #R250-01, Life Technologies) |
| Freeze Medium-DATA |
45% culture medium, 45% FBS (Cat. #10099-141, Gibco), 10% DMSO (Cat. #D2650, Sigma) |
Figure 1. AM (1-52)-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1 /Gα15/AMY2 and CHO-K1/Gα15 cells. The cells were loaded with Calcium-4 prior to stimulation with an AMY2 receptor agonist, AM (1-52). The intracellular calcium change was measured by FLIPR. Fluorescence signal were normalized and plotted against the log of the cumulative doses (5-fold dilution) of AM (1-52) (Mean ± SD, n = 4). The EC50 of AM (1-52) on this cells was 34 nM.
Notes:
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))
X is the logarithm of concentration. Y is the response
Y is RFU and starts at Bottom and goes to Top with a sigmoid shape.
2. Signal to background Ratio (S/B) = Top/Bottom
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