Journal
BIOPOLYMERS
Volume 99, Issue 5, Pages 314-325Publisher
WILEY
DOI: 10.1002/bip.22162
Keywords
PrRP; PrRP receptor; RF-amide peptide; NMR; chemical shifts; NOE; ROSETTA; ROSETTANMR; protein folding; GPCR; peptide receptor; structure/activity
Categories
Funding
- DFG [SFB 610, BE 1264-11]
- NIH [R01 MH090192, R01 GM GM080403]
- Office Of The Director
- Office Of Internatl Science &Engineering [1157751] Funding Source: National Science Foundation
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The prolactin releasing peptide (PrRP) is involved in regulating food intake and body weight homeostasis, but molecular details on the activation of the PrRP receptor remain unclear. C-terminal segments of PrRP with 20 (PrRP20) and 13 (PrRP8-20) amino acids, respectively, have been suggested to be fully active. The data presented herein indicate this is true for the wildtype receptor only; a 5-10-fold loss of activity was found for PrRP8-20 compared to PrRP20 at two extracellular loop mutants of the receptor. To gain insight into the secondary structure of PrRP, we used CD spectroscopy performed in TFE and SDS. Additionally, previously reported NMR data, combined with ROSETTANMR, were employed to determine the structure of amidated PrRP20. The structural ensemble agrees with the spectroscopic data for the full-length peptide, which exists in an equilibrium between - and 310-helix. We demonstrate that PrRP8-20's reduced propensity to form an -helix correlates with its reduced biological activity on mutant receptors. Further, distinct amino acid replacements in PrRP significantly decrease affinity and activity but have no influence on the secondary structure of the peptide. We conclude that formation of a primarily -helical C-terminal region of PrRP is critical for receptor activation. (c) 2012 Wiley Periodicals, Inc. Biopolymers 99: 273281, 2013.
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