A double headed serine proteinase inhibitor--human plasma kallikrein and elastase inhibitor--from Boophilus microplus larvae.
Preying on cattle, the hard tick Boophilus microplus causes heavy economical losses to Brazil. Tick proteins are a good target to be used as tools for tick control. Serine protease inhibitors from B. microplus larvae (BmTI) were preliminarily characterized. One-week-old larvae were the source of a 2% protein solution in 5 mM Tris-HCl, 20 mM NaCl, pH 7.4. The inhibitors were purified by affinity chromatography on trypsin-Sepharose, and ion-exchange chromatography on Resource Q column, and they separated in two major active peaks, corresponding to 10-kDa and 18-kDa proteins (BmTI-B and BmTI-A, respectively). Both purified proteins inhibited trypsin with Ki of 0.3 and 3.0 nM, respectively, but only the 18-kDa protein inhibited elastase (Ki 1.4 nM) and plasma kallikrein (Ki 120 nM). BmTI-A did not change prothrombin time (PT) and thrombin time (TT), but it increased activated partial thromboplastin time (APTT) was dose-dependent. The partial amino acid sequence indicated that BmTI-A belongs to the bovine pancreatic trypsin inhibitor (BPTI)-Kunitz type inhibitor family. These inhibitors (by their properties) play a role in the feeding process of the tick. Development of antibodies against these proteins may be used to impair the normal feeding and consequently, the parasite would be no longer viable.
Tanaka
A S
AS
Departamento de Bioquímica, UNIFESP-EPM, Sao Paulo, SP, Brazil. tanaka.bioq@epm.br
Andreotti
R
R
Gomes
A
A
Torquato
R J
RJ
Sampaio
M U
MU
Sampaio
C A
CA
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D006801:Humans; D007610:Kallikreins / Q000037:antagonists & inhibitors* / Q000097:blood*; D007814:Larva / Q000737:chemistry; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors*; D015842:Serine Proteinase Inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D013987:Ticks / Q000737:chemistry*
D015842:Serine Proteinase Inhibitors; D007610:Kallikreins; D010196:Pancreatic Elastase