Identification of extrinsic blood coagulation pathway inhibitors from the tick Ornithodoros savignyi (Acari: Argasidae).
The salt BaSO(4) selectively adsorbs two proteins from crude Ornithodoros savignyi salivary gland extract. They co-purify during reversed-phase HPLC, but can be separated by hydrophobic-interaction chromatography. Their molecular masses are 9333 and 9173Da. The 9.3kDa protein was designated BSAP1 and the 9.1kDa protein BSAP2. Their amino acid compositions show significant differences, in particular the presence of seven and eight cysteine residues in BSAP1 and BSAP2, respectively. The proteins do not contain gamma-carboxyglutamic acid, hydroxyproline, or hydroxylysine. The proteins do not inhibit the intrinsic coagulation cascade, but inhibit the extrinsic pathway. The observed inhibition is not due to inhibition of factor VII. Both proteins bind to membranes. BSAP1 binds neutral and negatively charged membranes more strongly than BSAP2. Its affinity for negative membranes is, however, much lower than for neutral membranes. In contrast, BSAP2 binds both membranes equally strongly. The binding of the proteins to the membranes was significantly lowered upon pre-incubation with Ca(2+).
Ehebauer
M T
MT
Department of Biochemistry, University of Pretoria, Pretoria, South Africa.
Mans
B J
BJ
Gaspar
A R M
AR
Neitz
A W H
AW
D000327:Adsorption; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification*; D001466:Barium Sulfate / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D008081:Liposomes; D008970:Molecular Weight; D026861:Ornithodoros / Q000737:chemistry*; D010314:Partial Thromboplastin Time; D011517:Prothrombin Time; D012469:Salivary Glands / Q000737:chemistry; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization
D000596:Amino Acids; D000925:Anticoagulants; D008081:Liposomes; D012471:Salivary Proteins and Peptides; D001466:Barium Sulfate