Stereochemical requirements for receptor recognition of the μ-opioid peptide endomorphin-1

M. Germana Paterlini, Francesca Avitabile, Beverly Gaul Ostrowski, David M Ferguson, Philip S Portoghese

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Abstract

A series of diastereoisomers of endomorphin-1 (EM1, Tyr1-Pro2-Trp3- Phe4-NH2) have been synthesized and their potency measured using the guinea pig ileum assay. [D-Phe4]EM1 possessed 1/10 the potency of EM1, while potencies of [D-Tyr1]EM1 and [D-Trp3]EM1 were 50- and 100-fold lower, respectively. Drastic loss of activity occurred in the [D-Pro2]EM1 peptide. The structural determinants for the inactivity and reduced potency of the diastereoisomers were investigated using NMR spectroscopy and conformational analysis. Simulations of trans-[D-Pro2]EM1 using NOE-derived distance constraints afforded well-defined structures in which Tyr and Trp side chains stack against the proline ring. The inactivity of [D- Pro2]EM1 was explained by structural comparison with EM1 (Podlogar et al., 1998, FEBS Lett. 439:13- 20). The two peptides showed an opposite orientation of the Trp3 residue with respect to Tyr1, thus suggesting a role of Pro2 as a stereochemical spacer in orienting Trp3 and Phe4 toward regions suitable for μ-receptor interaction. The agonist activity of [D-Tyr1]EM1 and [D-Trp3]EM1 was attributed to their ability to adopt low-energy conformations that mimic those of EM1. The requirements for μ-receptor activation were examined further by comparing EM1 with the μ-peptide [D-Ala2, MePhe4, Gly-ol]- enkephalin (DAMGO). Conformations of DAMGO with a Tyr -MePhe4 phenyl ring separation of ~12 Å, were found to mimic Tyr1-Phe4 of EM1, thus suggesting overlapping binding modes between these two peptides.

Original languageEnglish (US)
Pages (from-to)590-599
Number of pages10
JournalBiophysical journal
Volume78
Issue number2
DOIs
StatePublished - Jan 1 2000

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Opioid Peptides
Peptides
Enkephalins
Ileum
Proline
Guinea Pigs
Magnetic Resonance Spectroscopy
endomorphin 1

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Stereochemical requirements for receptor recognition of the μ-opioid peptide endomorphin-1. / Paterlini, M. Germana; Avitabile, Francesca; Ostrowski, Beverly Gaul; Ferguson, David M; Portoghese, Philip S.

In: Biophysical journal, Vol. 78, No. 2, 01.01.2000, p. 590-599.

Research output: Contribution to journalArticle

Paterlini, M. Germana ; Avitabile, Francesca ; Ostrowski, Beverly Gaul ; Ferguson, David M ; Portoghese, Philip S. / Stereochemical requirements for receptor recognition of the μ-opioid peptide endomorphin-1. In: Biophysical journal. 2000 ; Vol. 78, No. 2. pp. 590-599.
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abstract = "A series of diastereoisomers of endomorphin-1 (EM1, Tyr1-Pro2-Trp3- Phe4-NH2) have been synthesized and their potency measured using the guinea pig ileum assay. [D-Phe4]EM1 possessed 1/10 the potency of EM1, while potencies of [D-Tyr1]EM1 and [D-Trp3]EM1 were 50- and 100-fold lower, respectively. Drastic loss of activity occurred in the [D-Pro2]EM1 peptide. The structural determinants for the inactivity and reduced potency of the diastereoisomers were investigated using NMR spectroscopy and conformational analysis. Simulations of trans-[D-Pro2]EM1 using NOE-derived distance constraints afforded well-defined structures in which Tyr and Trp side chains stack against the proline ring. The inactivity of [D- Pro2]EM1 was explained by structural comparison with EM1 (Podlogar et al., 1998, FEBS Lett. 439:13- 20). The two peptides showed an opposite orientation of the Trp3 residue with respect to Tyr1, thus suggesting a role of Pro2 as a stereochemical spacer in orienting Trp3 and Phe4 toward regions suitable for μ-receptor interaction. The agonist activity of [D-Tyr1]EM1 and [D-Trp3]EM1 was attributed to their ability to adopt low-energy conformations that mimic those of EM1. The requirements for μ-receptor activation were examined further by comparing EM1 with the μ-peptide [D-Ala2, MePhe4, Gly-ol]- enkephalin (DAMGO). Conformations of DAMGO with a Tyr -MePhe4 phenyl ring separation of ~12 {\AA}, were found to mimic Tyr1-Phe4 of EM1, thus suggesting overlapping binding modes between these two peptides.",
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N2 - A series of diastereoisomers of endomorphin-1 (EM1, Tyr1-Pro2-Trp3- Phe4-NH2) have been synthesized and their potency measured using the guinea pig ileum assay. [D-Phe4]EM1 possessed 1/10 the potency of EM1, while potencies of [D-Tyr1]EM1 and [D-Trp3]EM1 were 50- and 100-fold lower, respectively. Drastic loss of activity occurred in the [D-Pro2]EM1 peptide. The structural determinants for the inactivity and reduced potency of the diastereoisomers were investigated using NMR spectroscopy and conformational analysis. Simulations of trans-[D-Pro2]EM1 using NOE-derived distance constraints afforded well-defined structures in which Tyr and Trp side chains stack against the proline ring. The inactivity of [D- Pro2]EM1 was explained by structural comparison with EM1 (Podlogar et al., 1998, FEBS Lett. 439:13- 20). The two peptides showed an opposite orientation of the Trp3 residue with respect to Tyr1, thus suggesting a role of Pro2 as a stereochemical spacer in orienting Trp3 and Phe4 toward regions suitable for μ-receptor interaction. The agonist activity of [D-Tyr1]EM1 and [D-Trp3]EM1 was attributed to their ability to adopt low-energy conformations that mimic those of EM1. The requirements for μ-receptor activation were examined further by comparing EM1 with the μ-peptide [D-Ala2, MePhe4, Gly-ol]- enkephalin (DAMGO). Conformations of DAMGO with a Tyr -MePhe4 phenyl ring separation of ~12 Å, were found to mimic Tyr1-Phe4 of EM1, thus suggesting overlapping binding modes between these two peptides.

AB - A series of diastereoisomers of endomorphin-1 (EM1, Tyr1-Pro2-Trp3- Phe4-NH2) have been synthesized and their potency measured using the guinea pig ileum assay. [D-Phe4]EM1 possessed 1/10 the potency of EM1, while potencies of [D-Tyr1]EM1 and [D-Trp3]EM1 were 50- and 100-fold lower, respectively. Drastic loss of activity occurred in the [D-Pro2]EM1 peptide. The structural determinants for the inactivity and reduced potency of the diastereoisomers were investigated using NMR spectroscopy and conformational analysis. Simulations of trans-[D-Pro2]EM1 using NOE-derived distance constraints afforded well-defined structures in which Tyr and Trp side chains stack against the proline ring. The inactivity of [D- Pro2]EM1 was explained by structural comparison with EM1 (Podlogar et al., 1998, FEBS Lett. 439:13- 20). The two peptides showed an opposite orientation of the Trp3 residue with respect to Tyr1, thus suggesting a role of Pro2 as a stereochemical spacer in orienting Trp3 and Phe4 toward regions suitable for μ-receptor interaction. The agonist activity of [D-Tyr1]EM1 and [D-Trp3]EM1 was attributed to their ability to adopt low-energy conformations that mimic those of EM1. The requirements for μ-receptor activation were examined further by comparing EM1 with the μ-peptide [D-Ala2, MePhe4, Gly-ol]- enkephalin (DAMGO). Conformations of DAMGO with a Tyr -MePhe4 phenyl ring separation of ~12 Å, were found to mimic Tyr1-Phe4 of EM1, thus suggesting overlapping binding modes between these two peptides.

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