ABSTRACT
We found that the tritium-labeled synthetic ACTH-like octapeptide leucocorticotropin corresponding to the 81-88 sequence of the precursor of human interleukin-1alpha ([3H]GKVLKKRR) is bound by the ACTH receptor of rat adrenal cortex with a high affinity and specificity (Kd 2.2 +/- 0.1 nM). This peptide was shown to exert no effect on the adenylate cyclase activity of the membranes of rat adrenal cortex in the concentration range from 1 to 1000 nM. Leucocorticotropin administration three times at doses of 10-20 microg/animal did not change the level of hydroxycorticosteroids (11-HOCS) in the rat adrenal glands in the absence of temperature action. At the same time, the peptide abolishes (at a dose of 20 microg/animal, three times) or significantly decreases (at a dose of 10 microg/animal, three times) the dramatic increase in the 11-HOCS content in the adrenal glands occurring in the case of cold or heat shock. Thus, leucocorticotropin normalizes the 11-HOCS level in the rat adrenal cortex during stress. The stress-protective effect of the peptide is mediated through the ACTH receptor.
Subject(s)
Adrenal Cortex Hormones/metabolism , Adrenal Cortex/drug effects , Interleukin-1alpha/pharmacology , Peptide Fragments/pharmacology , Protective Agents/pharmacology , Receptors, Corticotropin/agonists , Stress, Physiological/prevention & control , Administration, Intranasal , Adrenal Cortex/chemistry , Adrenal Cortex/metabolism , Adrenal Cortex Hormones/analysis , Adrenocorticotropic Hormone/chemistry , Amino Acid Sequence , Animals , Humans , Interleukin-1alpha/chemistry , Interleukin-1alpha/metabolism , Male , Molecular Sequence Data , Peptide Fragments/chemistry , Peptide Fragments/metabolism , Protective Agents/chemistry , Protective Agents/metabolism , Rats , Rats, Inbred Strains , Receptors, Corticotropin/metabolismABSTRACT
The effect of immunocortin, an ACTH-like decapeptide VKKPGSSVKV corresponding to the 11-20 sequence of the variable part of the human IgG1 heavy chain on the content of 11-hydroxycorticosteroids (CS) in rat adrenal glands and blood serum in vivo was studied. An intramuscular injection of immunocortin at a dose of 10 microg/kg was found in an hour to induce a twofold decrease in CS content in the adrenal glands and a 1.8-fold increase in the blood serum CS content. At the same time, an immunocortin dose of 100 microg/kg exerted practically no effect on the CS content and its dose of 1000 microg/kg increased the CS content both in adrenal glands and in blood serum by 1.6 and 2.2 times, respectively. Four hours after the injection of any of the three doses of immunocortin, the CS content in adrenal glands did not differ from the control value, and after 24 h the content decreased threefold. Immunocortin was shown to be bound by the ACTH receptors in the membranes of the rat adrenal cortex with a high affinity and specificity (inhibiting the specific binding of 125I-labeled ACTH-(11-24) peptide with Ki of 1.2 nM).
Subject(s)
11-Hydroxycorticosteroids/metabolism , Adrenal Cortex/metabolism , Adrenocorticotropic Hormone/chemistry , Immunoglobulin G/chemistry , Immunoglobulin G/pharmacology , Immunoglobulin Variable Region/chemistry , Peptide Fragments/pharmacology , 11-Hydroxycorticosteroids/blood , Adrenal Cortex/cytology , Adrenal Cortex/drug effects , Amino Acid Sequence , Animals , Cosyntropin/administration & dosage , Cosyntropin/pharmacology , Immunoglobulin G/administration & dosage , In Vitro Techniques , Iodine Radioisotopes , Male , Molecular Sequence Data , Peptide Fragments/administration & dosage , Peptide Fragments/chemistry , Rats , Rats, Wistar , Receptors, Corticotropin/metabolismABSTRACT
Adrenocorticotropic hormone (ACTH)-like peptide immunocortin (IMC) VKKPGSSVKV, corresponding to the amino acid sequence 11-20 of the variable part of human immunoglobulin G (IgG) 1 heavy chain, at concentrations of 10(-9)-10(-6) I was found to increase the adenylate cyclase activity in adrenal cortex membranes, while intramuscular injection of immunocortin at doses of 10-100 microg/kg was found to stimulate the secretion of 11-oxycorticosteroids (CS) from the adrenals to the bloodstream. Immunocortin was labeled with tritium to specific activity of 22 Ci/mmol. Receptor binding studies revealed that [(3)H]immunocortin ([(3)H]IMC) bound with high affinity and specificity to ACTH receptors on rat adrenal cortex membranes (K(d)=2.1+/-0.2 nM, B(max)=1.1+/-0.1 pmol/mg protein).