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Synthetic peptide KKRR corresponding to the human ACTH fragment 15–18 is an antagonist of the ACTH receptor

Abstract

Tritium-labeled synthetic fragments of human adrenocorticotropic hormone (ACTH) [3H]ACTH (11–24) and [3H]ACTH (15–18) with a specific activity of 22 and 26 Ci/mmol, respectively, were obtained. It was found that [3H]ACTH-(11–24) binds to membranes of the rat adrenal cortex with high affinity and high specificity (K d 1.8 ± 0.1 nM). Twenty nine fragments of ACTH (11–24) were synthesized, and their ability to inhibit the specific binding of [3H]ACTH (11–24) to adrenocortical membranes was investigated. The shortest active peptide was found to be an ACTH fragment (15–18) (KKRR) (K i 2.3 ± 0.2 nM), whose [3H] labeled derivative binds to rat adrenocortical membranes (K d 2.1 ± 0.1 nM) with a high affinity. The specific binding of [3H]ACTH-(15–18) was inhibited by 100% by unlabeled ACTH (11–24) (K i 2.0 ± 0.1 nM). ACTH (15–18) in the concentration range of 1–1000 nM did not affect the adenylate cyclase activity of adrenocortical membranes and, therefore, is an antagonist of the ACTH receptor.

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Abbreviations

ACTH:

adrenocorticotropic hormone

MC2R:

melanocortin-2 receptor

PAM:

phenacylamidomethyl

SEM:

standard error of the mean

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Correspondence to E. V. Navolotskaya.

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Original Russian Text © Yu.A. Kovalitskaya, A.A. Kolobov, E.A. Kampe-Nemm, Yu.A. Zolotarev, V.V. Yurovskii, V.B. Sadovnikov, V.M. Lipkin, E.V. Navolotskaya, 2008, published in Bioorganicheskaya Khimiya, 2008, Vol. 34, No. 1, pp. 29–35.

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Kovalitskaya, Y.A., Kolobov, A.A., Kampe-Nemm, E.A. et al. Synthetic peptide KKRR corresponding to the human ACTH fragment 15–18 is an antagonist of the ACTH receptor. Russ J Bioorg Chem 34, 24–29 (2008). https://doi.org/10.1134/S1068162008010020

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  • DOI: https://doi.org/10.1134/S1068162008010020

Key words

  • adrenocorticotropic hormone (ACTH)
  • peptides
  • receptors
  • adenylate cyclase
  • adrenal cortex