The production of recombinant Ornithodoros moubata Tick anticoagulant peptide involves isolating the target gene, which encodes the 1-60aa of Tick anticoagulant peptide. This gene is fused with an N-terminal 10xHis-SUMO-tag and C-terminal Myc-tag gene and then cloned into an appropriate expression vector. The constructed vector is introduced into E. coli cells via transformation. The E. coli cells express the recombinant protein upon certain induction and are lysed to release the expressed protein. Purification of the recombinant protein is typically achieved using affinity chromatography. The final step involves analyzing the protein's purity through SDS-PAGE. Its purity is over 85%.
Tick anticoagulant peptide (TAP) is a significant protein found in the saliva of the Ornithodoros moubata tick. This peptide, consisting of 60 amino acids, acts as a potent inhibitor of human blood coagulation factor Xa (fXa) [1]. Studies have shown that TAP is highly specific to factor Xa and is capable of binding to both an exosite and the active site of the enzyme [2]. Furthermore, TAP has been identified as a low molecular weight serine protease inhibitor, specifically targeting factor Xa [3][4]. The presence of TAP in the saliva of Ornithodoros moubata contributes to the tick's ability to feed on blood by inhibiting coagulation at the feeding site, allowing for a continuous blood meal [3].
References:
[1] S. Jordan, L. Waxman, D. Smith, & G. Vlasuk, Tick anticoagulant peptide: kinetic analysis of the recombinant inhibitor with blood coagulation factor x.alpha., Biochemistry, vol. 29, no. 50, p. 11095-11100, 1990. https://doi.org/10.1021/bi00502a012
[2] K. Bromfield, N. Quinsey, P. Duggan, & R. Pike, Approaches to selective peptidic inhibitors of factor xa, Chemical Biology & Drug Design, vol. 68, no. 1, p. 11-19, 2006. https://doi.org/10.1111/j.1747-0285.2006.00404.x
[3] A. Gaspar, A. Joubert, J. Crause, & A. Neitz, Isolation and characterization of an anticoagulant from the salivary glands of the tick, ornithodoros savignyi (acari: argasidae), Experimental and Applied Acarology, vol. 20, no. 10, p. 583-598, 1996. https://doi.org/10.1007/bf00052809
[4] A. Gaspar, J. Crause, & A. Neitz, Identification of anticoagulant activities in the salivary glands of the soft tick, ornithodoros savignyi, Experimental and Applied Acarology, vol. 19, no. 2, p. 117-127, 1995. https://doi.org/10.1007/bf00052551
