Bioactivity studies on atypical natural opioid hexapeptides processed from proenkephalin (PENK) precursor polypeptides

Engin Bojnik, Patrycja Kleczkowska, Ezequiel Marron Fernandez de Velasco, Maïthé Corbani, Fruzsina Babos, Andrzej W. Lipkowski, Anna Magyar, S. Benyhe

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Endogenous opioids are derived from four related polypeptide precursors: proenkephalin (PENK), prodynorphin (PDYN), pronociceptin (PNOC) and proopiomelanocortin (POMC). In mammals PENK encodes for four copy of Met-enkephalin, one octapeptide Met-enkephalin-Arg-Gly-Leu, one heptapeptide Met-enkephalin-Arg-Phe and a single copy of Leu-enkephalin. Our detailed bioinformatic search on the existing PENK sequences revealed several atypical hexapeptide Met-enkephalins in different vertebrate animals. They are located either in the second enkephalin unit or in the seventh enkephalin core position at the C-terminus. Altogether four different hexapeptide sequences were obtained representing eleven animal species: Met-enkephalin-Arg6 (YGGFMR) in the bird zebra finch, Met-enkephalin-Asp6 (YGGFMD), Met-enkephalin-Ile6 (YGGFMI) in zebrafish; and Met-enkephalin-Ser6 (YGGFMS) in two pufferfish species. All novel peptides were chemically synthesized and studied in receptor binding and G-protein activation assays performed on rat brain membranes. The four novel enkephalins were equipotent in stimulating G-proteins. Affinities of the peptides determined by equilibrium competition assays in receptor binding experiments were statistically different. At the MOP receptors the highest affinity (Ki 4nM) was obtained with the zebra finch peptide Met-enkephalin-Arg6. The pufferfish Met-enkephalin-Ser6 exhibited the highest affinity (Ki 6.7nM) at the DOP receptor. Phylogenetic neuropeptide libraries, defined here as a collection of mutationally different species variants of orthologous and paralogous peptide sequences, represent the natural molecular diversity of the neuropeptides. Such libraries can provide a wide range of structural information establishing comparative functional analyses. Since DNA sequencing data are rapidly increasing, more development in the natural peptide library concept is expected.

Original languageEnglish
Pages (from-to)29-35
Number of pages7
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume174
Issue number1
DOIs
Publication statusPublished - 2014

Fingerprint

Methionine Enkephalin
Bioactivity
Opioid Analgesics
Peptides
Enkephalins
Tetraodontiformes
Finches
Equidae
Arg(6)-Phe(7)-enkephalin-met
Neuropeptides
GTP-Binding Proteins
Assays
Animals
Leucine Enkephalin
proenkephalin
Peptide Library
Pro-Opiomelanocortin
Mammals
Birds
Zebrafish

Keywords

  • Enkephalin hexapeptides
  • Mu and delta opioid receptors
  • Natural chemical library
  • Proenkephalin evolution
  • Species differences

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Physiology
  • Medicine(all)

Cite this

Bioactivity studies on atypical natural opioid hexapeptides processed from proenkephalin (PENK) precursor polypeptides. / Bojnik, Engin; Kleczkowska, Patrycja; de Velasco, Ezequiel Marron Fernandez; Corbani, Maïthé; Babos, Fruzsina; Lipkowski, Andrzej W.; Magyar, Anna; Benyhe, S.

In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, Vol. 174, No. 1, 2014, p. 29-35.

Research output: Contribution to journalArticle

Bojnik, Engin ; Kleczkowska, Patrycja ; de Velasco, Ezequiel Marron Fernandez ; Corbani, Maïthé ; Babos, Fruzsina ; Lipkowski, Andrzej W. ; Magyar, Anna ; Benyhe, S. / Bioactivity studies on atypical natural opioid hexapeptides processed from proenkephalin (PENK) precursor polypeptides. In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology. 2014 ; Vol. 174, No. 1. pp. 29-35.
@article{9b1ba98034ed420e86a14ad3761779ee,
title = "Bioactivity studies on atypical natural opioid hexapeptides processed from proenkephalin (PENK) precursor polypeptides",
abstract = "Endogenous opioids are derived from four related polypeptide precursors: proenkephalin (PENK), prodynorphin (PDYN), pronociceptin (PNOC) and proopiomelanocortin (POMC). In mammals PENK encodes for four copy of Met-enkephalin, one octapeptide Met-enkephalin-Arg-Gly-Leu, one heptapeptide Met-enkephalin-Arg-Phe and a single copy of Leu-enkephalin. Our detailed bioinformatic search on the existing PENK sequences revealed several atypical hexapeptide Met-enkephalins in different vertebrate animals. They are located either in the second enkephalin unit or in the seventh enkephalin core position at the C-terminus. Altogether four different hexapeptide sequences were obtained representing eleven animal species: Met-enkephalin-Arg6 (YGGFMR) in the bird zebra finch, Met-enkephalin-Asp6 (YGGFMD), Met-enkephalin-Ile6 (YGGFMI) in zebrafish; and Met-enkephalin-Ser6 (YGGFMS) in two pufferfish species. All novel peptides were chemically synthesized and studied in receptor binding and G-protein activation assays performed on rat brain membranes. The four novel enkephalins were equipotent in stimulating G-proteins. Affinities of the peptides determined by equilibrium competition assays in receptor binding experiments were statistically different. At the MOP receptors the highest affinity (Ki 4nM) was obtained with the zebra finch peptide Met-enkephalin-Arg6. The pufferfish Met-enkephalin-Ser6 exhibited the highest affinity (Ki 6.7nM) at the DOP receptor. Phylogenetic neuropeptide libraries, defined here as a collection of mutationally different species variants of orthologous and paralogous peptide sequences, represent the natural molecular diversity of the neuropeptides. Such libraries can provide a wide range of structural information establishing comparative functional analyses. Since DNA sequencing data are rapidly increasing, more development in the natural peptide library concept is expected.",
keywords = "Enkephalin hexapeptides, Mu and delta opioid receptors, Natural chemical library, Proenkephalin evolution, Species differences",
author = "Engin Bojnik and Patrycja Kleczkowska and {de Velasco}, {Ezequiel Marron Fernandez} and Ma{\"i}th{\'e} Corbani and Fruzsina Babos and Lipkowski, {Andrzej W.} and Anna Magyar and S. Benyhe",
year = "2014",
doi = "10.1016/j.cbpb.2014.06.002",
language = "English",
volume = "174",
pages = "29--35",
journal = "Comparative biochemistry and physiology. B, Comparative biochemistry",
issn = "1096-4959",
publisher = "Elsevier Inc.",
number = "1",

}

TY - JOUR

T1 - Bioactivity studies on atypical natural opioid hexapeptides processed from proenkephalin (PENK) precursor polypeptides

AU - Bojnik, Engin

AU - Kleczkowska, Patrycja

AU - de Velasco, Ezequiel Marron Fernandez

AU - Corbani, Maïthé

AU - Babos, Fruzsina

AU - Lipkowski, Andrzej W.

AU - Magyar, Anna

AU - Benyhe, S.

PY - 2014

Y1 - 2014

N2 - Endogenous opioids are derived from four related polypeptide precursors: proenkephalin (PENK), prodynorphin (PDYN), pronociceptin (PNOC) and proopiomelanocortin (POMC). In mammals PENK encodes for four copy of Met-enkephalin, one octapeptide Met-enkephalin-Arg-Gly-Leu, one heptapeptide Met-enkephalin-Arg-Phe and a single copy of Leu-enkephalin. Our detailed bioinformatic search on the existing PENK sequences revealed several atypical hexapeptide Met-enkephalins in different vertebrate animals. They are located either in the second enkephalin unit or in the seventh enkephalin core position at the C-terminus. Altogether four different hexapeptide sequences were obtained representing eleven animal species: Met-enkephalin-Arg6 (YGGFMR) in the bird zebra finch, Met-enkephalin-Asp6 (YGGFMD), Met-enkephalin-Ile6 (YGGFMI) in zebrafish; and Met-enkephalin-Ser6 (YGGFMS) in two pufferfish species. All novel peptides were chemically synthesized and studied in receptor binding and G-protein activation assays performed on rat brain membranes. The four novel enkephalins were equipotent in stimulating G-proteins. Affinities of the peptides determined by equilibrium competition assays in receptor binding experiments were statistically different. At the MOP receptors the highest affinity (Ki 4nM) was obtained with the zebra finch peptide Met-enkephalin-Arg6. The pufferfish Met-enkephalin-Ser6 exhibited the highest affinity (Ki 6.7nM) at the DOP receptor. Phylogenetic neuropeptide libraries, defined here as a collection of mutationally different species variants of orthologous and paralogous peptide sequences, represent the natural molecular diversity of the neuropeptides. Such libraries can provide a wide range of structural information establishing comparative functional analyses. Since DNA sequencing data are rapidly increasing, more development in the natural peptide library concept is expected.

AB - Endogenous opioids are derived from four related polypeptide precursors: proenkephalin (PENK), prodynorphin (PDYN), pronociceptin (PNOC) and proopiomelanocortin (POMC). In mammals PENK encodes for four copy of Met-enkephalin, one octapeptide Met-enkephalin-Arg-Gly-Leu, one heptapeptide Met-enkephalin-Arg-Phe and a single copy of Leu-enkephalin. Our detailed bioinformatic search on the existing PENK sequences revealed several atypical hexapeptide Met-enkephalins in different vertebrate animals. They are located either in the second enkephalin unit or in the seventh enkephalin core position at the C-terminus. Altogether four different hexapeptide sequences were obtained representing eleven animal species: Met-enkephalin-Arg6 (YGGFMR) in the bird zebra finch, Met-enkephalin-Asp6 (YGGFMD), Met-enkephalin-Ile6 (YGGFMI) in zebrafish; and Met-enkephalin-Ser6 (YGGFMS) in two pufferfish species. All novel peptides were chemically synthesized and studied in receptor binding and G-protein activation assays performed on rat brain membranes. The four novel enkephalins were equipotent in stimulating G-proteins. Affinities of the peptides determined by equilibrium competition assays in receptor binding experiments were statistically different. At the MOP receptors the highest affinity (Ki 4nM) was obtained with the zebra finch peptide Met-enkephalin-Arg6. The pufferfish Met-enkephalin-Ser6 exhibited the highest affinity (Ki 6.7nM) at the DOP receptor. Phylogenetic neuropeptide libraries, defined here as a collection of mutationally different species variants of orthologous and paralogous peptide sequences, represent the natural molecular diversity of the neuropeptides. Such libraries can provide a wide range of structural information establishing comparative functional analyses. Since DNA sequencing data are rapidly increasing, more development in the natural peptide library concept is expected.

KW - Enkephalin hexapeptides

KW - Mu and delta opioid receptors

KW - Natural chemical library

KW - Proenkephalin evolution

KW - Species differences

UR - http://www.scopus.com/inward/record.url?scp=84903540253&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84903540253&partnerID=8YFLogxK

U2 - 10.1016/j.cbpb.2014.06.002

DO - 10.1016/j.cbpb.2014.06.002

M3 - Article

C2 - 24947211

AN - SCOPUS:84903540253

VL - 174

SP - 29

EP - 35

JO - Comparative biochemistry and physiology. B, Comparative biochemistry

JF - Comparative biochemistry and physiology. B, Comparative biochemistry

SN - 1096-4959

IS - 1

ER -