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Short peptide constructs mimic agonist sites of AT1R and BK receptors

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Abstract

Extracellular peptide ligand binding sites, which bind the N-termini of angiotensin II (AngII) and bradykinin (BK) peptides, are located on the N-terminal and extracellular loop 3 regions of the AT1R and BKRB1 or BKRB2 G-protein-coupled receptors (GPCRs). Here we synthesized peptides P15 and P13 corresponding to these receptor fragments and showed that only constructs in which these peptides were linked by S–S bond, and cyclized by closing the gap between them, could bind agonists. The formation of construct-agonist complexes was revealed by electron paramagnetic resonance spectra and fluorescence measurements of spin labeled biologically active analogs of AngII and BK (Toac1-AngII and Toac0-BK), where Toac is the amino acid-type paramagnetic and fluorescence quencher 2, 2, 6, 6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid. The inactive derivatives Toac3-AngII and Toac3-BK were used as controls. The interactions characterized by a significant immobilization of Toac and quenching of fluorescence in complexes between agonists and cyclic constructs were specific for each system of peptide-receptor construct assayed since no crossed reactions or reaction with inactive peptides could be detected. Similarities among AT, BKR, and chemokine receptors were identified, thus resulting in a configuration for AT1R and BKRB cyclic constructs based on the structure of the CXCR4, an α-chemokine GPCR-type receptor.

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Acknowledgments

This research was supported by Fundacao de Amparo à Pesquisa do Estado de Sao Paulo (FAPESP) and Conselho Nacional de Pesquisa Científica e Tecnológica (CNPq). CRN, SS, and SIS are CNPq’ research fellows, RFFV is recipient of FAPESP pos-doctoral fellowship, and DDL is recipient of CNPq PhD fellowship.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biophysics, Federal University of Sao Paulo, Sao Paulo, CEP 04044-020, Brazil

    Douglas D. Lopes, Renata F. F. Vieira, Luciana Malavolta, Erick F. Poletti, Suma I. Shimuta, Antonio C. M. Paiva, Laerte Oliveira & Clovis R. Nakaie

  2. Department of Physiological Science, Faculdade de Ciências Médicas da Santa Casa de Sao Paulo, Sao Paulo, CEP 01221-020, Brazil

    Luciana Malavolta

  3. Department of Biochemistry Institute of Chemistry, University of São Paulo, Sao Paulo, CEP 05513-970, Brazil

    Shirley Schreier

Authors
  1. Douglas D. Lopes
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  2. Renata F. F. Vieira
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  3. Luciana Malavolta
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  4. Erick F. Poletti
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  5. Suma I. Shimuta
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  6. Antonio C. M. Paiva
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  7. Shirley Schreier
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  8. Laerte Oliveira
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  9. Clovis R. Nakaie
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Corresponding author

Correspondence to Clovis R. Nakaie.

Additional information

Antonio C. M. Paiva: Deceased.

D. D. Lopes, R. F. F. Vieira, and L. Malavolta contributed equally to the manuscript.

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Lopes, D.D., Vieira, R.F.F., Malavolta, L. et al. Short peptide constructs mimic agonist sites of AT1R and BK receptors. Amino Acids 44, 835–846 (2013). https://doi.org/10.1007/s00726-012-1405-9

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  • DOI: https://doi.org/10.1007/s00726-012-1405-9

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