Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein
Open Access
- 25 June 2020
- Vol. 9 (6), 513
- https://doi.org/10.3390/pathogens9060513
Abstract
A synthetic 20 amino acid peptide of the ribosomal protein P0 from ticks, when conjugated to keyhole limpet hemocyanin from Megathura crenulata and used as an immunogen against Rhipicephalus microplus and Rhipicephalus sanguineus s.l. species, has shown efficacies of around 90%. There is also experimental evidence of a high efficacy of this conjugate against Amblyomma mixtum and Ixodes ricinus species, which suggest that this antigen could be a good broad-spectrum anti-tick vaccine candidate. In this study, the P0 peptide (pP0) was chemically conjugated to Bm86 as a carrier protein. SDS-PAGE analysis of this conjugate demonstrated that it is highly heterogeneous in size, carrying from 1 to 18 molecules of pP0 per molecule of Bm86. Forty-nine out of the 54 lysine residues and the N-terminal end of Bm86 were found partially linked to pP0 by using LC-MS/MS analysis and the combination of four different softwares. Several post-translational modifications of Bm86 protein were also identified by mass spectrometry. High immunogenicity and efficacy were achieved when dogs and cattle were vaccinated with the pP0–Bm86 conjugate and challenged with R. sanguineus s.l. and R. microplus, respectively. These results encourage the development of this antigen with promising possibilities as an anti-tick vaccine.This publication has 64 references indexed in Scilit:
- Dosimetry of N6-Formyllysine Adducts Following [13C2H2]-Formaldehyde Exposures in RatsChemical Research in Toxicology, 2013
- Quantitative Analysis of Histone Modifications: Formaldehyde Is a Source of Pathological N6-Formyllysine That Is Refractory to Histone DeacetylasesPLoS Genetics, 2013
- Identification of cross-linked peptides from complex samplesNature Methods, 2012
- Bm86 homologues and novel ATAQ proteins with multiple epidermal growth factor (EGF)-like domains from hard and soft ticksInternational Journal for Parasitology, 2010
- Two initial vaccinations with the Bm86-based Gavacplus vaccine against Rhipicephalus (Boophilus) microplusinduce similar reproductive suppression to three initial vaccinations under production conditionsBMC Veterinary Research, 2010
- Finding Chimeras: a Bioinformatics Strategy for Identification of Cross-linked PeptidesMolecular & Cellular Proteomics, 2010
- Identification and characterization of Rhipicephalus (Boophilus) microplus candidate protective antigens for the control of cattle tick infestationsZeitschrift für Parasitenkunde, 2009
- Formaldehyde and leukemia: Epidemiology, potential mechanisms, and implications for risk assessmentEnvironmental and Molecular Mutagenesis, 2009
- Detergent binding explains anomalous SDS-PAGE migration of membrane proteinsProceedings of the National Academy of Sciences of the United States of America, 2009
- Gene silencing of the tick protective antigens, Bm86, Bm91 and subolesin, in the one-host tick Boophilus microplus by RNA interferenceInternational Journal for Parasitology, 2007