Bowman-Birk Protease Inhibitor from Vigna unguiculata Seeds Enhances the Action of Bradykinin-Related Peptides

Molecules

Alice da Cunha M. Álvares²  Elisabeth Ferroni Schwartz³  Nathalia Oda Amaral⁴  Neidiane Rosa Trindade⁴  Gustavo Rodrigues Pedrino⁴  Luciano Paulino Silva¹  Sonia Maria de Freitas² 

[1]Embrapa Recursos Genéticos e Biotecnologia, Laboratório de Espectrometria de Massa, PBI, Parque Estação Biológica, Final W5 Norte, Asa Norte, Brasília, DF 70770-917, Brazil

[2] E-Mail:

[3]Laboratório de Biofísica, Instituto de Biologia, Departamento de Biologia Celular, Universidade de Brasília-UnB, Quadra 604, Asa Norte, Bloco J 1° andar, Brasília, DF 70910-900, Brazil

[4] E-Mail:

[5]Laboratório de Toxinologia, Departamento de Ciências Fisiológicas, Universidade de Brasília-UnB, Quadra 604, Asa Norte, Bloco J Térreo, Brasília, DF 70910-900, Brazil

[6] E-Mail:

[7]Center of Neuroscience and Cardiovascular Physiology, Department of Physiological Sciences, Biological Sciences Institute, Federal University of Goiás, Estrada do Campus, Goiânia, GO 74690-900, Brazil

[8] E-Mails:

关键词: arterial blood pressure;    bradykinin;    Bowman-Birk inhibitor;    circular dichroism;    dynamic light scattering;    fluorescence spectroscopy;   

DOI  :  10.3390/molecules191117536

来源: mdpi

【 摘 要 】

The hydrolysis of bradykinin (Bk) by different classes of proteases in plasma and tissues leads to a decrease in its half-life. Here, Bk actions on smooth muscle and in vivo cardiovascular assays in association with a protease inhibitor, Black eyed-pea trypsin and chymotrypsin inhibitor (BTCI) and also under the effect of trypsin and chymotrypsin were evaluated. Two synthetic Bk-related peptides, Bk₁ and Bk₂, were used to investigate the importance of additional C-terminal amino acid residues on serine protease activity. BTCI forms complexes with Bk and analogues at pH 5.0, 7.4 and 9.0, presenting binding constants ranging from 10³ to 10⁴ M⁻¹. Formation of BTCI-Bk complexes is probably driven by hydrophobic forces, coupled with slight conformational changes in BTCI. In vitro assays using guinea pig (Cavia porcellus) ileum showed that Bk retains the ability to induce smooth muscle contraction in the presence of BTCI. Moreover, no alteration in the inhibitory activity of BTCI in complex with Bk and analogous was observed. When the BTCI and BTCI-Bk complexes were tested in vivo, a decrease of vascular resistance and consequent hypotension and potentiating renal and aortic vasodilatation induced by Bk and Bk₂ infusions was observed. These results indicate that BTCI-Bk complexes may be a reliable strategy to act as a carrier and protective approach for Bk-related peptides against plasma serine proteases cleavage, leading to an increase in their half-life. These findings also indicate that BTCI could remain stable in some tissues to inhibit chymotrypsin or trypsin-like enzymes that cleave and inactivate bradykinin in situ.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.