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Synthesis, receptor binding, and crosslinking of photoactive analogues of neuropeptide Y.
Journal article

Synthesis, receptor binding, and crosslinking of photoactive analogues of neuropeptide Y.

  • 1995-01-01
Published in:
  • Journal of receptor and signal transduction research. - 1995
Affinity Labels
Amino Acid Sequence
Animals
Azirines
Binding Sites
Cell Line
Cross-Linking Reagents
In Vitro Techniques
Molecular Sequence Data
Neuropeptide Y
Phenylalanine
Photochemistry
Receptors, Neuropeptide Y
Swine
English Five photoactive analogues of porcine neuropeptide Y (NPY), a 36 amino acid hormone of the pancreatic polypeptide family, have been synthesized by solid phase peptide synthesis method, Fmoc/tBu strategy and carefully characterized. The analogues contain the photoactivatable amino acid 4'-(3-trifluoromethyl)-3H-diazirine-3-yl-phenyl-alanine ((Tmd)Phe) individually at different positions (1, 20, 21, 27 or 36) instead of tyrosine in the wildtype sequence. Affinity to membranes prepared from SMS-KAN cells, which stably express the Y2 receptor has been investigated by measuring the displacement of 125I-Bolton Hunter-NPY. After incubation of the membranes with different concentrations of the crosslinker and subsequent photolysis, the specific binding of 125I-Bolton Hunter-NPY at those membranes was tested in order to quantify the crosslinking efficiency. Whereas [(Tmd)Phe20] NPY, [(Tmd)Phe21] NPY and [(Tmd)Phe27] NPY revealed highest affinity to the Y2 receptor, crosslinking was most efficient when Tyr36 was replaced by (Tmd)Phe. This is in good agreement with the previously suggested C-terminal binding site of neuropeptide Y.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/112935
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