title: Self-assembled protein arrays from an Ornithodoros moubata salivary gland expression library.
abstract: Protein interactions play a critical role in the regulation of many biological events and their study in a high-throughput format has become a key area of proteomic research. Nucleid Acid Programmable Protein Arrays (NAPPA) technology allows the construction of protein arrays from cDNA expression libraries in high-throughput cell-free systems to study protein interaction and functions. Tick saliva contains antihemostatic, anti-inflammatory, and immunosuppressive proteins that counteract the host hemostatic, immune, and inflammatory responses allowing the ingestion of host blood and facilitating its infection by the tick-borne pathogens. Identification of such proteins and their functions could help in the selection of antigenic targets for the development of antitick and transmission-blocking vaccines. With that aim, we have prepared a cDNA expression library from the salivary glands of Ornithodoros moubata and subsequently produced a self-assembled protein microarray using 480 randomly selected clones from that library. The reproducibility of the array, its representativeness of the tick salivary protein repertoire, and the functionality of the in situ expressed proteins have been checked, demonstrating that it is a suitable tool for the identification and functional characterization of soft tick salivary molecules that interact with host proteins. Several clones in the array were shown to bind to human recombinant P-selectin. One of them was a likely secreted tick phospholipase A2, which may represent a potential new ligand for P-selectin. As these salivary molecules are likely involved in blood meal acquisition through the modulation of the host immune and hemostatic responses, this new high-throughput tool could open new avenues for development of new therapeutic agents and control strategies against ticks and tick-borne pathogens.
authors.affiliation: Parasitología Animal, Instituto de Recursos Naturales y Agrobiología de Salamanca (IRNASA, CSIC), Cordel de Merinas, 40-52, 37008 Salamanca, Spain.
authors.forename: Raul
authors.initials: R
authors.lastname: Manzano-Román
authors.forename: Veronica
authors.initials: V
authors.lastname: Díaz-Martín
authors.forename: Maria
authors.initials: M
authors.lastname: González-González
authors.forename: Sergio
authors.initials: S
authors.lastname: Matarraz
authors.forename: Angel Francisco
authors.initials: AF
authors.lastname: Álvarez-Prado
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
authors.forename: Alberto
authors.initials: A
authors.lastname: Orfao
authors.forename: Ricardo
authors.initials: R
authors.lastname: Pérez-Sánchez
authors.forename: Manuel
authors.initials: M
authors.lastname: Fuentes
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000032:analysis* / Q000378:metabolism; D002474:Cell-Free System; D005260:Female; D015723:Gene Library; D057166:High-Throughput Screening Assays; D006801:Humans; D008297:Male; D026861:Ornithodoros / Q000235:genetics / Q000378:metabolism*; D019007:P-Selectin / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods; D011994:Recombinant Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012469:Salivary Glands / Q000166:cytology / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D060829:Arthropod Proteins; D019007:P-Selectin; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides