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The synthetic peptide octarphin (TPLVTLFK) corresponding to the sequence 12-19 of β-endorphin, a selective agonist of nonopioid β-endorphin receptor, was labeled with tritium to a specific activity of 29 Ci/mmol. [(3)H]Octarphin was found to bind to high-affinity naloxone-insensitive binding sites on membranes isolated from rat adrenal cortex (K(d) = 35.7 ± 2.3 nM, B(max) = 41.0 ± 3.6 pmol/mg protein). The binding specificity study revealed that these binding sites were insensitive not only to naloxone but to α-endorphin, γ-endorphin, [Met(5) ]enkephalin, and [Leu(5) ]enkephalin as well. At the same time, the [(3) H]octarphin-specific binding with adrenal cortex membranes was inhibited by unlabeled β-endorphin (K(i) = 32.9 ± 3.8 nM). Octarphin at concentrations of 10(-9) -10(-6) M was found to inhibit the adenylate cyclase activity in adrenocortical membranes, whereas intranasal injection of octarphin at doses of 5 and 20 µg/rat was found to reduce the secretion of corticosterone from the adrenals to the bloodstream. Thus, octarphin decreases the adrenal cortex functional activity through the high affinity binding to nonopioid receptor of β-endorphin., (Copyright © 2012 European Peptide Society and John Wiley & Sons, Ltd.)

Two selective agonists of nonopioid β-endorphin receptor, synthetic peptides TPLVTLFK (octarphin) and SLTCLVKGFY (immunorphin), were labeled with tritium to specific activity of 29 and 25 Ci/mmol, respectively. Both labeled peptides were found to bind to high-affinity naloxone-insensitive binding sites on the membranes isolated from the rat myocardium (Kd = 2.0 ± 0.2 and 2.5 ± 0.3 nM, respectively). The [(3)H]octarphin specific binding to the myocardial membranes was inhibited by unlabeled β-endorphin (Ki = 1.9 ± 0.2 nM) and immunorphin (Ki = 2.2 ± 0.3 nM). The [(3)H]immunorphin specific binding with the membranes was inhibited by unlabeled β-endorphin (Ki = 2.3 ± 0.3 nM) and octarphin (Ki = 2.4 ± 0.3 nM). The binding specificity study revealed that these binding sites were insensitive not only to naloxone but also to α-endorphin, γ-endorphin, [Met(5)]enkephalin and [Leu(5)]enkephalin. Thus, β-endorphin, immunorphin and octarphin bind to the common high-affinity naloxone-insensitive receptor of the rat myocardial membranes., (Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd.)

The synthetic peptide TPLVTLFK corresponding to the sequence 12-19 of beta-endorphin (referred to as octarphin) was found to bind to high-affinity naloxone-insensitive binding sites on membranes isolated from the rat brain cortex (K(d) = 2.6 +/- 0.2 nM). The binding specificity study revealed that these binding sites were insensitive not only to naloxone but also to alpha-endorphin, gamma-endorphin, [Met(5)]enkephalin, and [Leu(5)]enkephalin, as well. The [(3)H]octarphin specific binding with brain membranes was inhibited by unlabeled beta-endorphin (K(i) = 2.4 +/- 0.2 nM) and a selective agonist of nonopioid beta-endorphin receptor decapeptide immunorphin SLTCLVKGFY (K(i) = 2.9 +/- 0.2 nM). At the same time, unlabeled octarphin completely (by 100%) inhibited the specific binding of [(3)H]immunorphin with membranes (K(i) = 2.8 +/- 0.2 nM). Thus, octarphin binds with a high affinity and specificity to nonopioid receptor of beta-endorphin on rat brain cortex membranes., ((c) 2010 European Peptide Society and John Wiley & Sons, Ltd.)

Selective agonist of nonopioid beta-endorphin receptor decapeptide immunorphin (SLTCLVKGFY) was labeled with tritium (the specific activity of 24 Ci/mmol). [3H]Immunorphin was found to bind to nonopioid beta-endorphin receptor of mouse peritoneal macrophages (Kd = 2.0 +/- 0.1 nM). The [3H]immunorphin specific binding with macrophages was inhibited by unlabeled beta-endorphin (Ki = 2.9 +/- 0.2 nM) and was not inhibited by unlabeled naloxone, alpha-endorphin, gamma-endorphin and [Met5]enkephalin (Ki > 10 microM). Thirty fragments of beta-endorphin have been synthesized and their ability to inhibit the [3H]immunorphin specific binding to macrophages was studied. Unlabeled fragment 12-19 (TPLVTLFK, the author's name of the peptide octarphin) was found to be the shortest peptide possessing practically the same inhibitory activity as beta-endorphin (Ki = 3.1 +/- 0.3 nM). The peptide octarphin was labeled with tritium (the specific activity of 28 Ci/mmol). [3H]Octarphin was found to bind to macrophages with high affinity (Kd = 2.3 +/- 0.2 nM). The specific binding of [3H]octarphin was inhibited by unlabeled immunorphin and beta-endorphin (Ki = 2.4 +/- 0.2 and 2.7 +/- 0.2 nM, respectively)., (Copyright 2008 European Peptide Society and John Wiley & Sons, Ltd.)

beta-Endorphin-like decapeptide immunorphin (SLTCLVKGFY), a selective agonist of non-opioid beta-endorphin receptor, was labeled with tritium to specific activity of 24 Ci/mmol. It was used for the detection and characterization of non-opioid beta-endorphin receptors on rat adrenal cortex membranes (Kd1 = 39.6 +/- 2.0 nM, Bmax1 = 40.7 +/- 2.3 pmol/mg protein; Kd2 = 0.25 +/- 0.01 micro M, Bmax2 = 187.8 +/- 9.4 pmol/mg protein). beta-Endorphin was found to inhibit the [3H]immunorphin specific binding to membranes (Ki = 70.0 +/- 9.2 nM); naloxone, [Met5]enkephalin, and alpha- and gamma-endorphins tested in parallel were inactive. Immunorphin at concentrations of 10(-9)-10(-6) M was found to inhibit the adenylate cyclase activity in adrenocortical membranes, while intramuscular injection of immunorphin at doses of 10-100 micro g/kg was found to reduce the secretion of 11-oxycorticosteroids from the adrenals to the bloodstream., (Copyright 2004 MAIK)

Tritium-labeled selective agonist of non-opioid beta-endorphin receptor, the decapeptide immunorphine ([3H]SLTCLVKGFY) with specific activity of 24 Ci/mmol has been prepared. By its use, non-opioid beta-endorphin receptors were revealed and characterized on mouse peritoneal macrophages and rat myocardium, spleen, adrenal, and brain membranes. The non-opioid beta-endorphin receptor of macrophages has in addition to immunorphine (Kd of the [3H]immunorphine-receptor complex was 2.4 +/- 0.1 nM) and beta-endorphin (Ki of the [3H]immunorphine specific binding was 2.9 +/- 0.2 nM) a high affinity for Fc-fragment of human IgG1, pentarphine (VKGFY), cyclopentarphine [cyclo(VKGFY)], and [Pro3]pentarphine (VKPFY) (Ki values were 0.0060 +/- 0.0004, 2.7 +/- 0.2, 2.6 +/- 0.2, and 2.8 +/- 0.2 nM, respectively) and is insensitive to naloxone and [Met5]enkephalin (Ki > 100 microM). Treatment of macrophages with trypsin resulted in the loss of their ability for the specific binding of [3H]immunorphine. Values of the specific binding of 8.4 nM [3H]immunorphine to rat adrenal, spleen, myocardium, and brain membranes were determined to be 1146.0 +/- 44.7, 698.6 +/- 28.1, 279.1 +/- 15.4, and 172.2 +/- 1.8 fmol/mg protein, respectively. Unlabeled beta-endorphin, pentarphine, [Pro3]pentarphine, cyclopentarphine, cyclodipentarphine [cyclo(VKGFYVKGFY)], and Fc-fragment of IgG1 inhibited the binding of [3H]immunorphine to membranes from these organs. No specific binding of [3H]immunorphine to rat liver, lung, kidney, and intestine membranes was found.

Beta-endorphin-like peptide immunorphin (SLTCLVKGFY), a selective agonist of nonopioid beta-endorphin receptor, was labeled with tritium to specific activity of 24 Ci/mmol. It was used for the detection and characterization of nonopioid beta-endorphin receptors on rat adrenal cortex membranes (Kd = 31.6 +/- 0.2 nM, Bmax = 37.4 +/- 2.2 pmol/mg protein). Immunorphin at concentrations of 10(-9) to 10(-6) M was found to inhibit the adenylate cyclase activity in adrenal cortex membranes, while intramuscular injection of immunorphin at doses of 10-100 microg/kg was found to reduce the secretion of 11-oxycorticosteroids from the adrenals to the bloodstream.

It was shown that beta-endorphin and the synthetic decapeptide SLTCLVKGFY that corresponds to the amino acid sequence 364-373 of the human IgG heavy chain (referred to as immunorphin) is able to stimulate growth of the human T-lymphoblastoid cell line Jurkat. The antagonist of opioid receptors naloxone did not inhibit the stimulating effect of the peptides. Studies on [(3)H]-immunorphin binding to Jurkat cell receptors have demonstrated that it binds with high affinity to naloxone-insensitive receptors (K(d) = 1.3 nM; n = 5.2 x 10(5)). Unlabeled beta-endorphin and the 6-10 fragment of immunorphin completely inhibited the labeled ligand specific binding to naloxone-insensitive receptors on T lymphocytes (K(i) = 1.4 x 10(-7) and 3.7 x 10(-5) M, respectively). Thus, beta-endorphin and immunorphin share the naloxone-insensitive receptors on human T-lymphoblastoid cell line Jurkat.