Self-assembled protein arrays from an ornithodoros moubata salivary gland expression library

Raul Manzano-Román, Veronica Díaz-Martín, Maria González-González, Sergio Matarraz, Angel Francisco Álvarez-Prado, Joshua LaBaer, Alberto Orfao, Ricardo Pérez-Sánchez, Manuel Fuentes

Research output: Contribution to journalArticlepeer-review

31 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)5972-5982
Number of pages11
JournalJournal of Proteome Research
Issue number12
StatePublished - Dec 7 2012


  • expression libraries
  • interactions
  • protein arrays
  • salivary glands
  • ticks

ASJC Scopus subject areas

  • Biochemistry
  • General Chemistry


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