TY - JOUR
T1 - Release of immunomodulatory peptides at bacterial membrane interfaces as a novel strategy to fight microorganisms
AU - Viana de freitas, Thiago
AU - Karmakar, Utsa
AU - Vasconcelos, Andreanneg.
AU - Santos, Michelea.
AU - Oliveira do vale lira, Bianca
AU - Costa, Samuelribeiro
AU - Barbosa, Ederalves
AU - Cardozo-Fh, José
AU - Correa, Rafael
AU - Ribeiro, Dalilaj.s.
AU - Prates, Mauravianna
AU - Magalhães, Kellyg.
AU - Soller ramada, Marcelohenrique
AU - Roberto de souza almeida leite, José
AU - Bloch, Carlos
AU - Lima de oliveira, Aline
AU - Vendrell, Marc
AU - Brand, Guilhermedotto
N1 - Funding Information:
Authors would also like to thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES, www.capes.gov.br/pt/ ) - Finance Code 001; G.D.B. acknowledges funds from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, www.cnpq.br/ ) grant 407515/2021-6; Fundação de amparo a pesquisa do DF (FAP-DF, www.fap.df.gov.br/ ) grants 0193.000866/2015 and 0193.001566/2017. M. V. acknowledges funds from an ERC CoG ( 771443 , DYNAFLUORS).
Funding Information:
The authors wish to express their gratitude to Laboratório de Neurofarmacologia, Neuropharma, Instituto de Biologia, Universidade de Brasília, more specifically, to Adolfo Carlos Barros de Souza. Also, we are grateful to Embrapa Recursos Genéticos e Biotecnologia, Brasília, DF, Brazil, and the Analytic Central of the Institute of Chemistry of the University of Brasília, for the support in infrastructure. T. V. d. F. U. K. A. G. V. M. A. S. B. O. d. V. L. S. R. C. E. A. B. J. C.-Fh. R. C. D. J. S. R. and M. V. P. investigation; T. V. F. U. K. K. G. M. M. H. S. R. J. R. d. S. A. L. C. B. Jr, A. L. d. O. P. M. V. and G. D. B. methodology; T. V. d. F. and U. K. conceptualization; K. G. M. M. H. S. R. J. R. d. S. A. L. C. B. Jr, A. L. d. O. M. V. and G. D. B. formal analysis; M. V. and G. D. B. supervision; M. V. and G. D. B. writing–original draft. Authors would also like to thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES, www.capes.gov.br/pt/) - Finance Code 001; G.D.B. acknowledges funds from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, www.cnpq.br/) grant 407515/2021-6; Fundação de amparo a pesquisa do DF (FAP-DF, www.fap.df.gov.br/) grants 0193.000866/2015 and 0193.001566/2017. M. V. acknowledges funds from an ERC CoG (771443, DYNAFLUORS).
Publisher Copyright:
© 2023 The Authors
PY - 2023/2/21
Y1 - 2023/2/21
N2 - Cationic and amphiphilic peptides can be used as homing devices to accumulate conjugated antibiotics to bacteria-enriched sites and promote efficient microbial killing. However, just as important as tackling bacterial infections, is the modulation of the immune response in this complex microenvironment. In the present report, we designed a peptide chimaera called Chim2, formed by a membrane-active module, an enzyme hydrolysis site, and a formyl peptide receptor 2 (FPR2) agonist. This molecule was designed to adsorb onto bacterial membranes, promote their lysis, and upon hydrolysis by local enzymes, release the FPR2 agonist sequence for activation and recruitment of immune cells. We synthesized the isolated peptide modules of Chim2 and characterized their biological activities independently and as a single polypeptide chain. We conducted antimicrobial assays, along with other tests aiming at the analyses of the cellular and immunological responses. In addition, assays using vesicles as models of eukaryotic and prokaryotic membranes were conducted, and solution structures of Chim2 were generated by 1H NMR. Chim2 is antimicrobial, adsorbs preferentially to negatively charged vesicles while adopting an α-helix structure, and exposes its disorganized tail to the solvent, which facilitates hydrolysis by tryptase-like enzymes, allowing the release of the FPR2 agonist fragment. This fragment was shown to induce accumulation of the cellular activation marker, lipid bodies, in mouse macrophages and the release of immunomodulatory interleukins. In conclusion, these data demonstrate that peptides with antimicrobial and immunomodulatory activities can be considered for further development as drugs.
AB - Cationic and amphiphilic peptides can be used as homing devices to accumulate conjugated antibiotics to bacteria-enriched sites and promote efficient microbial killing. However, just as important as tackling bacterial infections, is the modulation of the immune response in this complex microenvironment. In the present report, we designed a peptide chimaera called Chim2, formed by a membrane-active module, an enzyme hydrolysis site, and a formyl peptide receptor 2 (FPR2) agonist. This molecule was designed to adsorb onto bacterial membranes, promote their lysis, and upon hydrolysis by local enzymes, release the FPR2 agonist sequence for activation and recruitment of immune cells. We synthesized the isolated peptide modules of Chim2 and characterized their biological activities independently and as a single polypeptide chain. We conducted antimicrobial assays, along with other tests aiming at the analyses of the cellular and immunological responses. In addition, assays using vesicles as models of eukaryotic and prokaryotic membranes were conducted, and solution structures of Chim2 were generated by 1H NMR. Chim2 is antimicrobial, adsorbs preferentially to negatively charged vesicles while adopting an α-helix structure, and exposes its disorganized tail to the solvent, which facilitates hydrolysis by tryptase-like enzymes, allowing the release of the FPR2 agonist fragment. This fragment was shown to induce accumulation of the cellular activation marker, lipid bodies, in mouse macrophages and the release of immunomodulatory interleukins. In conclusion, these data demonstrate that peptides with antimicrobial and immunomodulatory activities can be considered for further development as drugs.
KW - antibiotics
KW - antimicrobial peptide (AMP)
KW - drug design
KW - immunology
KW - inflammation
KW - nuclear magnetic resonance (NMR)
KW - pattern recognition receptor (PRR)
KW - peptide chemical synthesis
U2 - 10.1016/j.jbc.2023.103056
DO - 10.1016/j.jbc.2023.103056
M3 - Article
C2 - 36822328
SN - 0021-9258
VL - 299
SP - 103056
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 4
M1 - 103056
ER -