title: Longipin: An Amyloid Antimicrobial Peptide from the Harvestman Acutisoma longipes (Arachnida: Opiliones) with Preferential Affinity for Anionic Vesicles.
abstract: In contrast to vertebrate immune systems, invertebrates lack an adaptive response and rely solely on innate immunity in which antimicrobial peptides (AMPs) play an essential role. Most of them are membrane active molecules that are typically unstructured in solution and adopt secondary/tertiary structures upon binding to phospholipid bilayers. This work presents the first characterization of a constitutive AMP from the hemolymph of an Opiliones order animal: the harvestman Acutisoma longipes. This peptide was named longipin. It presents 18 aminoacid residues (SGYLPGKEYVYKYKGKVF) and a positive net charge at neutral pH. No similarity with other AMPs was observed. However, high sequence similarity with heme-lipoproteins from ticks suggested that longipin might be a protein fragment. The synthetic peptide showed enhanced antifungal activity against Candida guilliermondii and C. tropicalis yeasts (MIC: 3.8-7.5 μM) and did not interfered with VERO cells line viability at all concentrations tested (200-0.1 μM). This selectivity against microbial cells is related to the highest affinity of longipin for anionic charged vesicles (POPG:POPC) compared to zwitterionic ones (POPC), once microbial plasma membrane are generally more negatively charged compared to mammalian cells membrane. Dye leakage from carboxyfluorescein-loaded POPG:POPC vesicles suggested that longipin is a membrane active antimicrobial peptide and FT-IR spectroscopy showed that the peptide chain is mainly unstructured in solution or in the presence of POPC vesicles. However, upon binding to POPG:POPC vesicles, the FT-IR spectrum showed bands related to β-sheet and amyloid-like fibril conformations in agreement with thioflavin-T binding assays, indicating that longipin is an amyloid antimicrobial peptide.
authors.affiliation: Programa Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.|Laboratório Especial de Toxinologia Aplicada, Instituto Butantan, São Paulo, Brazil.
authors.forename: Raphael Santa Rosa
authors.identifier: http://orcid.org/0000-0002-6461-6627
authors.initials: RS
authors.lastname: Sayegh
authors.affiliation: Unidade de Sequenciamento de Proteínas e Peptídeos, Instituto Butantan, São Paulo, Brazil.|Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, Brazil.
authors.forename: Isabel de Fátima Correia
authors.initials: IF
authors.lastname: Batista
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Instituto Butantan, São Paulo, Brazil.
authors.forename: Robson Lopes de
authors.initials: RL
authors.lastname: Melo
authors.affiliation: Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Karin A
authors.initials: KA
authors.lastname: Riske
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Sirlei
authors.initials: S
authors.lastname: Daffre
authors.affiliation: Centro de Investigaciones en Quimica Biológica de Córdoba (CIQUIBIC, UNC-CONICET), Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba, Argentina.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Montich
authors.affiliation: Programa Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.|Laboratório Especial de Toxinologia Aplicada, Instituto Butantan, São Paulo, Brazil.
authors.forename: Pedro Ismael
authors.initials: PI
authors.lastname: da Silva Junior
mesh_terms: D000682:Amyloid / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D001097:Arachnida / Q000737:chemistry / Q000235:genetics; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D001419:Bacteria / Q000254:growth & development*; D002175:Candida / Q000254:growth & development*; D002522:Chlorocebus aethiops; D014709:Vero Cells
chemical_list: D000682:Amyloid; D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins