Synthesis of enzymatically resistant nociceptin-related peptides containing a carbamic acid residue

Melinda Ligeti, Sz. Bősze, A. Csámpai, Özge Gündüz, M. Al-Khrasani, A. Rónai, Hedvig Medzihradszky-Schweiger, S. Benyhe, A. Borsodi, F. Hudecz, Anna Magyar

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Nociceptin, a 17-amino acid peptide (FGGFTGARKSARKLANQ, N/OFQ), is the endogenous ligand of the nociceptin/orphanin FQ (NOP) receptor. This receptor-ligand system is involved in various physiological as well as pathophysiological mechanisms, but owing to the peptidic structure, it is rapidly degraded by enzymes. The enzymatic digestion of nociceptin involves mainly aminopeptidases and yields Noc(2-17)-OH and other smaller fragments. We aimed at increasing the enzymatic stability against aminopeptidases in the case of peptide Noc(1-13)-NH2, which possesses the minimum sequence capable of interacting with the NOP receptor. Therefore we developed a new procedure for the synthesis of peptides with the carbamic acid residue [...-NH-CH(R)-CO-NH-CO-NH-CH(Q)-CO-...). A set of four carbamic acid-nociceptin derivatives were produced. The carbamic acid residue was incorporated into the inner part of the peptides, building on solid phase, by using a suitable dipeptide fragment with carbamic acid residue produced by a simple and efficient three-step solution phase procedure. Enzymatic stability of carbamic acid peptides was studied in the presence of aminopeptidase M (AP-M) and in rat brain membrane homogenate. The receptor-binding properties were also studied by radioligand binding assay on crude rat brain membranes and the activity of the ligands were analyzed on isolated mouse vas deferens (MVD) tissues. We found that incorporation of the carbamic acid residue into the N-terminal part of nociceptin significantly increases the resistance against AP-M. We observed the decrease of binding affinities to the NOP receptor in case of the peptides modified in the N-terminal portion. Consequently, the incorporation of the carbamic acid residue into peptides can be proposed as a promising and reasonable tool for increasing enzymatic stability, where the native molecule is less sensitive for carbamic acid residue-related structural change.

Original languageEnglish
Pages (from-to)481-490
Number of pages10
JournalJournal of Peptide Science
Volume12
Issue number7
DOIs
Publication statusPublished - Jul 2006

Fingerprint

Peptides
Carbon Monoxide
Aminopeptidases
Ligands
Rats
Brain
Membranes
CD13 Antigens
Radioligand Assay
Vas Deferens
carbamic acid
nociceptin
Dipeptides
Digestion
Assays
Tissue
Derivatives
Amino Acids
Molecules
Enzymes

Keywords

  • Aminopeptidase M
  • Carbamic acid residue
  • Enzymatic stability
  • GTPγS assay
  • Mouse vas deferens bioassay
  • Nociceptin
  • NOP receptor
  • Radioligand binding
  • Rat brain membrane homogenate

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Synthesis of enzymatically resistant nociceptin-related peptides containing a carbamic acid residue. / Ligeti, Melinda; Bősze, Sz.; Csámpai, A.; Gündüz, Özge; Al-Khrasani, M.; Rónai, A.; Medzihradszky-Schweiger, Hedvig; Benyhe, S.; Borsodi, A.; Hudecz, F.; Magyar, Anna.

In: Journal of Peptide Science, Vol. 12, No. 7, 07.2006, p. 481-490.

Research output: Contribution to journalArticle

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AU - Ligeti, Melinda

AU - Bősze, Sz.

AU - Csámpai, A.

AU - Gündüz, Özge

AU - Al-Khrasani, M.

AU - Rónai, A.

AU - Medzihradszky-Schweiger, Hedvig

AU - Benyhe, S.

AU - Borsodi, A.

AU - Hudecz, F.

AU - Magyar, Anna

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KW - GTPγS assay

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KW - Radioligand binding

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