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Insights into cardiovascular effects of proline-rich oligopeptide (Bj-PRO-10c) revealed by structure–activity analyses: dissociation of antihypertensive and bradycardic effects

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Abstract

We have previously reported that the proline-rich decapeptide from Bothrops jararaca (Bj-PRO-10c) causes potent and sustained antihypertensive and bradycardic effects in SHR. These activities are independent of ACE inhibition. In the present study, we used the Ala-scan approach to evaluate the importance of each amino acid within the sequence of Bj-PRO-10c (Pyr1-Asn2-Trp3-Pro4-His5-Pro6-Gln7-Ile8-Pro9-Pro10). The antihypertensive and bradycardic effects of the analogues Bj-PRO-10c Ala3, Bj-PRO-10c Ala7, Bj-PRO-10c Ala8 were similar to those of Bj-PRO-10c, whereas the analogues Bj-PRO-10c Ala2, Bj-PRO-10c Ala4, Bj-PRO-10c Ala5, Bj-PRO-10c Ala9, and Bj-PRO-10c Ala10 kept the antihypertensive activity and lost bradycardic activity considerably. In contrast, Bj-PRO-10c Ala1 and Bj-PRO-10c Ala6 were unable to provoke any cardiovascular activity. In summary, we demonstrated that (1) the Pyr1 and Pro6 residues are essential for both, the antihypertensive and bradycardic effects of Bj-PRO-10c; (2) Ala-scan approach allowed dissociating blood pressure reduction and bradycardic effects. Conformational properties of the peptides were examined by means of circular dichroism (CD) spectroscopy. The different Ala-scan analogues caused either an increase or decrease in the type II polyproline helix content compared to Bj-PRO-10c. The complete loss of activity of the Pro6 → Ala6 mutant is probably due to the fact that in the parent peptide the His5-Pro6 bond can exist in the cis configuration, which could correspond to the conformation of this bond in the bound state. Current data support the Bj-PRO-10c as a promising leader prototype to develop new agents to treat cardiovascular diseases and its co-morbidities.

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Abbreviations

ACE:

Angiotensin-I converting enzyme

Ala-scan:

Alanine scan

AsS:

Argininosuccinate synthetase

Bj :

Bothrops jararaca

Bj-PRO:

Bothrops jararaca-proline-rich oligopeptides

CD:

Circular dichroism

HR:

Heart rate

i.v.:

Intravenous

MAP:

Mean arterial pressure

MDLA:

Alpha-methyl-dl-aspartic acid

NO:

Nitric oxide

PAP:

Pulsatile arterial pressure

Pyr:

Pyroglutamic acid

SHR:

Spontaneously hypertensive rat

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Acknowledgments

The majority of the in vivo study was used as part of the requirements for a master’s degree by J.F.B. Paschoal. This research was supported by Grants provided by Fundação de Amparo à Pesquisa do Estado de São Paulo (CAT/Cepid-FAPESP, 98/14307-9), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Edital Toxinologia—n. 63/2010, AUXPE 1593/2011. G.P.B.C. is recipient of CNPq PhD fellowship. S.S. is recipient of CNPq research fellowship. The authors would like to acknowledge Beatriz L. Fernandes for critical review, Maria José da Silva and Isaías França da Silva for secretarial assistance, and Vera Pontieri for technical assistance.

Confict of interest

The authors declare that they have no conflict of interest.

Author Contributions

Participated in research design: A.C.M. Camargo and D. Ianzer;

Conducted experiments: J.F.B. Paschoal, J. Yamaguchi, J.R.R. Miranda, C.H. Xavier, G. Carretero and D. Ianzer;

Contributed new reagents or analytic tools: R.L. Melo, R.A.S. Santos and S. Schreier;

Performed data analysis: D. Ianzer, C.H. Xavier, G. Carretero and S. Schreier;

Wrote or contributed to the writing of the manuscript: C.H. Xavier, G. Carretero, S. Schreier, A.C.M. Camargo, and D. Ianzer.

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Authors and Affiliations

  1. Special Laboratory of Applied Toxinology—CAT/Cepid, Butantan Institute, Av. Vital Brasil, 1500, Sao Paulo, SP, 05503-900, Brazil

    Juliana F. B. Paschoal, Juliana Yamaguchi, José R. R. Miranda, Robson L. Melo, Antonio C. M. Camargo & Danielle Ianzer

  2. Department of Biochemistry, Federal University of Sao Paulo (UNIFESP), Sao Paulo, SP, Brazil

    Juliana F. B. Paschoal, Juliana Yamaguchi & José R. R. Miranda

  3. Department of Biochemistry, Institute of Chemistry, University of Sao Paulo (USP), Sao Paulo, SP, Brazil

    Gustavo Carretero & Shirley Schreier

  4. Laboratory of Hypertension, Federal University of Minas Gerais (UFMG), Belo Horizonte, MG, Brazil

    Robson A. S. Santos & Danielle Ianzer

  5. Department of Physiological Sciences, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil

    Carlos H. Xavier

Authors
  1. Juliana F. B. Paschoal
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  9. Antonio C. M. Camargo
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Correspondence to Danielle Ianzer.

Additional information

J. F. B. Paschoal and J. Yamaguchi should be considered as co-first authors.

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Paschoal, J.F.B., Yamaguchi, J., Miranda, J.R.R. et al. Insights into cardiovascular effects of proline-rich oligopeptide (Bj-PRO-10c) revealed by structure–activity analyses: dissociation of antihypertensive and bradycardic effects. Amino Acids 46, 401–413 (2014). https://doi.org/10.1007/s00726-013-1630-x

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