Structural Studies of a Lipid-Binding Peptide from Tunicate Hemocytes with Anti-Biofilm Activity.
Clavanins is a class of peptides (23aa) histidine-rich, free of post-translational modifications. Clavanins have been studied largely for their ability to disrupt bacterial membranes. In the present study, the interaction of clavanin A with membranes was assessed by dynamic light scattering, zeta potential and permeabilization assays. We observed through those assays that clavanin A lysis bacterial cells at concentrations corresponding to its MIC. Further, the structure and function of clavanin A was investigated. To better understand how clavanin interacted with bacteria, its NMR structure was elucidated. The solution state NMR structure of clavanin A in the presence of TFE-d3 indicated an α-helical conformation. Secondary structures, based on circular dichroism measurements in anionic sodium dodecyl sulfate (SDS) and TFE (2,2,2-trifluorethanol), in silico lipid-peptide docking and molecular simulations with lipids DPPC and DOPC revealed that clavanin A can adopt a variety of folds, possibly influencing its different functions. Microcalorimetry assays revealed that clavanin A was capable of discriminating between different lipids. Finally, clavanin A was found to eradicate bacterial biofilms representing a previously unrecognized function.
Departamento Biologia, Instituto de Ciências Biológicas, Programa de pós-graduação em Genética e Biotecnologia, Universidade Federal de Juiz de Fora, Juiz de Fora-MG, 36036-900, Brazil.|SInova, Pós-graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, MS, Brazil.
Osmar N
ON
Silva
Laboratório de RMN, Instituto de Química, Universidade Federal de Goiás, C.P. 131, 74001-970 Goiânia, GO, Brazil.
Eliane S F
ES
Alves
Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada.
César
C
de la Fuente-Núñez
SInova, Pós-graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, MS, Brazil.
Suzana M
SM
Ribeiro
Central Research Facility, Department of Chemistry and Department of Biotechnology, Indian Institute of Technology Kharagpur, India.
Santi M
SM
Mandal
Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028 Lisbon, Portugal.
Diana
D
Gaspar
Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028 Lisbon, Portugal.
Ana S
AS
Veiga
Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028 Lisbon, Portugal.
Miguel A R B
MA
Castanho
Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Cesar A S
CA
Andrade
Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Jessica M
JM
Nascimento
SInova, Pós-graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, MS, Brazil.
Isabel C M
IC
Fensterseifer
SInova, Pós-graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, MS, Brazil.
William F
WF
Porto
Department of Cellular Biology, Institute of Biological Sciences, University of Brasilia, Brasilia, 70910900, Brazil.
Jose R
JR
Correa
Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada.
Robert E W
RE
Hancock
Central Research Facility, Department of Chemistry and Department of Biotechnology, Indian Institute of Technology Kharagpur, India.
Suresh
S
Korpole
Laboratório de RMN, Instituto de Química, Universidade Federal de Goiás, C.P. 131, 74001-970 Goiânia, GO, Brazil.|Instituto de Química, Universidade de Brasília, C.P. 04478, 70910-000, Brasília, DF, Brazil.
Aline L
AL
Oliveira
Laboratório de RMN, Instituto de Química, Universidade Federal de Goiás, C.P. 131, 74001-970 Goiânia, GO, Brazil.
Luciano M
LM
Liao
Departamento Biologia, Instituto de Ciências Biológicas, Programa de pós-graduação em Genética e Biotecnologia, Universidade Federal de Juiz de Fora, Juiz de Fora-MG, 36036-900, Brazil.|SInova, Pós-graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, MS, Brazil.|Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Centro de Análises Proteômicas e Bioquímicas, Universidade Católica de Brasília, Brazil.
Octavio L
OL
Franco
D000818:Animals; D001419:Bacteria / Q000187:drug effects*; D018407:Bacterial Physiological Phenomena / Q000187:drug effects; D018441:Biofilms / Q000187:drug effects*; D001798:Blood Proteins / Q000737:chemistry* / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects; D002942:Circular Dichroism; D000067493:Dynamic Light Scattering; D006434:Hemocytes / Q000737:chemistry / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D062105:Molecular Docking Simulation; D017433:Protein Structure, Secondary; D014561:Urochordata / Q000737:chemistry / Q000378:metabolism*
D001798:Blood Proteins; D008051:Lipid Bilayers; C106713:clavanin A