PMID: 11567096

 

    Legend: Gene, Sites

Title : Functional and protein chemical characterization of the N-terminal domain of the rat corticotropin-releasing factor receptor 1

Abstract :
  1. Rat corticotropin-releasing factor receptor 1 ( rCRFR1 ) was produced either in transfected HEK 293 cells as a complex glycosylated protein or in the presence of the mannosidase I inhibitor kifunensine as a high mannose glycosylated protein
  2. The altered glycosylation did not influence the biological function of rCRFR1 as demonstrated by competitive binding of rat urocortin ( rUcn ) or human/rat corticotropin-releasing factor (h/ rCRF ) and agonist-induced cAMP accumulation
  3. The low production rate of the N-terminal domain of rCRFR1 ( rCRFR1-NT) by transfected HEK 293 cells, was increased by a factor of 100 in the presence of kifunensine
  4. The product, rCRFR1-NT-Kif, bound rUcn specifically (K(D) = 27 nM) and astressin (K(I) = 60 nM)
  5. This affinity was 10-fold lower than the affinity of full length rCRFR1
  6. However, it was sufficiently high for rCRFR1-NT-Kif to serve as a model for the N-terminal domain of rCRFR1
  7. With protein fragmentation , Edman degradation, and mass spectrometric analysis, evidence was found for the signal peptide cleavage site C-terminally to Thr(23) and three disulfide bridges between precursor residues 30 and 54, 44 and 87, and 68 and 102
  8. Of all putative N-glycosylation sites in positions 32, 38, 45, 78, 90, and 98, all Asn residues except for Asn(32) were glycosylated to a significant extent
  9. No O-glycosylation was observed
Output (sent_index, trigger, protein, sugar, site):
  • 1. corticotropin-releasing, , corticotropin-releasing factor receptor 1, a high mannose glycosylated protein, -
  • 1. factor, , corticotropin-releasing factor receptor 1, a high mannose glycosylated protein, -
  • 1. glycosylated, , corticotropin-releasing factor receptor 1, a high mannose glycosylated protein, -
  • 1. glycosylated, , protein, -, -
  • 8. Asn, , -, all Asn residues, Asn residues
  • 8. glycosylated, , -, -, Asn residues
  • 8. sites, , -, positions 32, 38, 45, 78, 90, and 98, positions 32, 38, 45, 78, 90, and 98,
Output(Part-Of) (sent_index, protein, site):
  • 0. corticotropin-releasing factor receptor 1, domain
  • 3. rCRFR1, domain
  • 6. rCRFR1, domain
*Output_Site_Fusion* (sent_index, protein, sugar, site):

 

 

Protein NCBI ID SENTENCE INDEX
urocortin 29151 2
rCRFR1 58959 1,2,3,4,5,6
rCRF 81648 2
corticotropin-releasing factor 81648 2