Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein

Overview

Journal Pathogens

Date 2020 Jul 8

PMID 32630414

Citations 14

Authors

Alina Rodriguez Mallon

Luis Javier Gonzalez

Pedro Enrique Encinosa Guzman

Gervasio Henrique Bechara

Gustavo Seron Sanches

Satomy Pousa

Gleysin Cabrera

Ania Cabrales

Hilda Garay

Raul Mejias

Jose Raul Lopez Alvarez

Yamil Bello Soto

Fabiola Almeida

Osmany Guirola

Rafmary Rodriguez Fernandez

Alier Fuentes Castillo

Luis Mendez

Samanta Jimenez

Alexei Licea-Navarro

Madelon Portela

Rosario Duran

Mario Pablo Estrada

Affiliations

Soon will be listed here.

Abstract

A synthetic 20 amino acid peptide of the ribosomal protein P0 from ticks, when conjugated to keyhole limpet hemocyanin from and used as an immunogen against and s.l. species, has shown efficacies of around 90%. There is also experimental evidence of a high efficacy of this conjugate against and 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 s.l. and , respectively. These results encourage the development of this antigen with promising possibilities as an anti-tick vaccine.

Citing Articles

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References

Radulovic Z, Kim T, Porter L, Sze S, Lewis L, Mulenga A . A 24-48 h fed Amblyomma americanum tick saliva immuno-proteome. BMC Genomics. 2014; 15:518. PMC: 4099483. DOI: 10.1186/1471-2164-15-518. View

Fields G, NOBLE R . Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids. Int J Pept Protein Res. 1990; 35(3):161-214. DOI: 10.1111/j.1399-3011.1990.tb00939.x. View

Smith P, Krohn R, Hermanson G, Mallia A, Gartner F, Provenzano M . Measurement of protein using bicinchoninic acid. Anal Biochem. 1985; 150(1):76-85. DOI: 10.1016/0003-2697(85)90442-7. View

Tabor A, Ali A, Rehman G, Rocha Garcia G, Zangirolamo A, Malardo T . Cattle Tick -Host Interface: A Review of Resistant and Susceptible Host Responses. Front Cell Infect Microbiol. 2018; 7:506. PMC: 5732177. DOI: 10.3389/fcimb.2017.00506. View

Rodriguez-Mallon A, Encinosa P, Mendez-Perez L, Bello Y, Fernandez R, Garay H . High efficacy of a 20 amino acid peptide of the acidic ribosomal protein P0 against the cattle tick, Rhipicephalus microplus. Ticks Tick Borne Dis. 2015; 6(4):530-7. DOI: 10.1016/j.ttbdis.2015.04.007. View

Chu F, Baker P, Burlingame A, Chalkley R . Finding chimeras: a bioinformatics strategy for identification of cross-linked peptides. Mol Cell Proteomics. 2009; 9(1):25-31. PMC: 2808265. DOI: 10.1074/mcp.M800555-MCP200. View

de Oliveira P, Bechara G, Denardi S, Nunes E, Mathias M . Morphological characterization of the ovary and oocytes vitellogenesis of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae). Exp Parasitol. 2005; 110(2):146-56. DOI: 10.1016/j.exppara.2004.12.016. View

Nijhof A, Balk J, Postigo M, Rhebergen A, Taoufik A, Jongejan F . Bm86 homologues and novel ATAQ proteins with multiple epidermal growth factor (EGF)-like domains from hard and soft ticks. Int J Parasitol. 2010; 40(14):1587-97. PMC: 2998001. DOI: 10.1016/j.ijpara.2010.06.003. View

Perez-Perez D, Bechara G, Machado R, Andrade G, Del Vecchio R, Pedroso M . Efficacy of the Bm86 antigen against immature instars and adults of the dog tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae). Vet Parasitol. 2009; 167(2-4):321-6. DOI: 10.1016/j.vetpar.2009.09.034. View

10.

Schilling B, Row R, Gibson B, Guo X, Young M . MS2Assign, automated assignment and nomenclature of tandem mass spectra of chemically crosslinked peptides. J Am Soc Mass Spectrom. 2003; 14(8):834-50. DOI: 10.1016/S1044-0305(03)00327-1. View

11.

Almazan C, Moreno-Cantu O, Moreno-Cid J, Galindo R, Canales M, Villar M . Control of tick infestations in cattle vaccinated with bacterial membranes containing surface-exposed tick protective antigens. Vaccine. 2011; 30(2):265-72. DOI: 10.1016/j.vaccine.2011.10.102. View

12.

Almazan C, Lagunes R, Villar M, Canales M, Rosario-Cruz R, Jongejan F . Identification and characterization of Rhipicephalus (Boophilus) microplus candidate protective antigens for the control of cattle tick infestations. Parasitol Res. 2009; 106(2):471-9. PMC: 2797406. DOI: 10.1007/s00436-009-1689-1. View

13.

Rodriguez M, Rubiera R, Penichet M, Montesinos R, Cremata J, Falcon V . High level expression of the B. microplus Bm86 antigen in the yeast Pichia pastoris forming highly immunogenic particles for cattle. J Biotechnol. 1994; 33(2):135-46. DOI: 10.1016/0168-1656(94)90106-6. View

14.

Gotze M, Pettelkau J, Schaks S, Bosse K, Ihling C, Krauth F . StavroX--a software for analyzing crosslinked products in protein interaction studies. J Am Soc Mass Spectrom. 2011; 23(1):76-87. DOI: 10.1007/s13361-011-0261-2. View

15.

Canales M, Enriquez A, Ramos E, Cabrera D, Dandie H, Soto A . Large-scale production in Pichia pastoris of the recombinant vaccine Gavac against cattle tick. Vaccine. 1997; 15(4):414-22. DOI: 10.1016/s0264-410x(96)00192-2. View

16.

Parizi L, Reck Jr J, Oldiges D, Guizzo M, Seixas A, Logullo C . Multi-antigenic vaccine against the cattle tick Rhipicephalus (Boophilus) microplus: a field evaluation. Vaccine. 2012; 30(48):6912-7. DOI: 10.1016/j.vaccine.2012.08.078. View

17.

Lu S, Fan S, Yang B, Li Y, Meng J, Wu L . Mapping native disulfide bonds at a proteome scale. Nat Methods. 2015; 12(4):329-31. DOI: 10.1038/nmeth.3283. View

18.

Cabrera G, Salazar V, Montesino R, Tambara Y, Struwe W, Leon E . Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). Glycobiology. 2015; 26(3):230-50. DOI: 10.1093/glycob/cwv096. View

19.

Rath A, Glibowicka M, Nadeau V, Chen G, Deber C . Detergent binding explains anomalous SDS-PAGE migration of membrane proteins. Proc Natl Acad Sci U S A. 2009; 106(6):1760-5. PMC: 2644111. DOI: 10.1073/pnas.0813167106. View

20.

de la Fuente J, Almazan C, Canales M, Perez de la Lastra J, Kocan K, Willadsen P . A ten-year review of commercial vaccine performance for control of tick infestations on cattle. Anim Health Res Rev. 2007; 8(1):23-8. DOI: 10.1017/S1466252307001193. View