Structural properties of orexins for activation of their receptors.
The closely related neuropeptides orexin A and orexin B mediate their actions, including the regulation of sleep and appetite, by the activation of the orexin 1 and 2 receptors. To elucidate the structural prerequisites for receptor activation and subtype selectivity, we performed multiple amino acid substitutions within the sequence of orexin A and human orexin B-(6-28)-peptide and analyzed their solution structures by CD spectroscopy and their activity at both receptors in Ca(2+) mobilization assays. For orexin A, we showed that the basic amino acids within the segment of residues 6-14 were important for the activation of both receptors. Furthermore, we showed that the restriction via disulfide bonds is not required to maintain the active structure of orexin A. The kink region of h orexin B has been shown to be important for Ox(2)R selectivity, which is not mediated by the restriction of the turn structure. Additionally, we showed that no particular secondary structure is required for receptor subtype selectivity.
Lang
Manja
M
Institute of Biochemistry, University of Leipzig, Germany.
Bufe
Bernd
B
De Pol
Silvia
S
Reiser
Oliver
O
Meyerhof
Wolfgang
W
Beck-Sickinger
Annette G
AG
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D002942:Circular Dichroism; D018076:DNA, Complementary; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009479:Neuropeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D064446:Orexin Receptors; D000068797:Orexins; D010455:Peptides / Q000138:chemical synthesis / Q000235:genetics; D043562:Receptors, G-Protein-Coupled; D018013:Receptors, Neuropeptide / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
D018076:DNA, Complementary; C000596229:HCRT protein, human; D047908:Intracellular Signaling Peptides and Proteins; D009479:Neuropeptides; D064446:Orexin Receptors; D000068797:Orexins; D010455:Peptides; D043562:Receptors, G-Protein-Coupled; D018013:Receptors, Neuropeptide