A critical role for the short intracellular C terminus in receptor activity-modifying protein function

Receptor activity-modifying proteins (RAMPs) interact with and modify the behavior of the calcitonin receptor (CTR) and calcitonin receptor-like receptor (CLR). We have examined the contribution of the short intracellular C terminus, using constructs that delete the last eight amino acids of each RAMP. C-Terminal deletion of individual RAMPs had little effect on the signaling profile induced when complexed with CLR in COS-7 or human embryonic kidney (HEK)293 cells. Likewise, confocal microscopy revealed each of the mutant RAMPs translocated hemagglutinin-tagged CLR to the cell surface. In contrast, a pronounced effect of RAMP C-terminal truncation was seen for RAMP/CTRa complexes, studied in COS-7 cells, with significant attenuation of amylin receptor phenotype induction that was stronger for RAMP1 and -2 than RAMP3. The loss of amylin binding upon C-terminal deletion could be partially recovered with overexpression of Gαs, suggesting an impact of the RAMP C terminus on coupling of G proteins to the receptor complex. In HEK293 cells the c-Myc-RAMP1 C-terminal deletion mutant showed high receptor-independent cell surface expression; however, this construct showed low cell surface expression when expressed alone in COS-7 cells, indicating interaction of RAMPs with other cellular components via the C terminus. This mutant also had reduced cell surface expression when coexpressed with CTR. Thus, this study reveals important functionality of the RAMP C-terminal domain and identifies key differences in the role of the RAMP C terminus for CTR versus CLR-based receptors.

The training load of aerial skiing

This study quantified the training load experienced by elite aerial skiers. Nine elite female aerial skiers were monitored during 16 training sessions over a 13 day period. Time-motion, landing impact and heart rate (HR) data were measured from 688 jumps using integrated GPS, accelerometer and HR transmitters while rating of perceived exertion (RPE) was taken using Borg's scale. Each jump was delineated into five components from the GPS time-motion data to determine the work to rest ratios. Participants completed 16 ± 3 jumps per session with a work to rest ratio of 1.9:1 Heart rates averaged 65 ± 3.1% HRmax and peaked at 85 ± 4.4% HRmax while and an RPE score of 12 ± 1 was evoked. Landing impacts were significantly higher (p ≤ 0.001) when participants jumped off ramps with a larger take-off angle or when they completed jumps with a mid-air rotation. The training load experienced by elite aerial skiers may be causative of the high incidence of injuries reported. Significantly differing levels of impact load during the study suggest training load for these athletes can be easily modified and periodised allowing optimised performance and minimised injury.