Purification and partial characterization of kininogenase activity from Schistosoma mansoni adult worms.
An enzyme presenting kallikrein-like activity (designated sK1) was purified from the supernatant of Schistosoma mansoni adult worm homogenate. The enzyme cleaves bradykinin from purified rat plasma kininogen. Activity was optimal at pH 9.0 and the enzyme showed amidolytic activity, since it hydrolysed the kallikrein synthetic substrate D-Pro-Phe-Arg-p-nitroanilide. The activity of sK1 upon rat plasma kininogen was strongly inhibited by the serine proteinase inhibitors phenylmethanesulfonyl fluoride, aprotinin or soybean trypsin inhibitor, but not by ethylenediaminetetraacetic acid or sodium tetrathionate. The molecular mass of sK1, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, was 66 kDa and the pI value, estimated by analytical chromatofocusing, was 4.2. Physical and chemical properties suggest that sK1 is a serine proteinase of the kallikrein family. Evidence is presented which suggests that sK1 is a component of the tegumental surface of the parasite and the levels of its activity in the male adult worm are approximately 21 times higher than those in the female adult worm. The intravenous injection of 3 micrograms of sK1 into an anaesthetized rat induced a drastic reduction in the arterial blood pressure of the animal. This effect lasted for about 1 min, and was followed by a progressive recovery of the arterial pressure. Neither bradycardia nor cardiac arrhythmias were noticed, suggesting a peripheral vasodilation effect. The presence of sK1 on the surface of adult male worms could play an important role in the wandering capacity of coupled worms into the visceral vasculature of the host.
Carvalho
W S
WS
Department of Social Pharmacy, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, Brazil.
Lopes
C T
CT
Juliano
L
L
Coelho
P M
PM
Cunha-Melo
J R
JR
Beraldo
W T
WT
Pesquero
J L
JL
D000818:Animals; D007611:Aprotinin / Q000737:chemistry; D001794:Blood Pressure / Q000502:physiology; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D004492:Edetic Acid / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007610:Kallikreins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007704:Kininogens / Q000097:blood; D008297:Male; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D012550:Schistosoma mansoni / Q000737:chemistry / Q000201:enzymology*; D013776:Tetrathionic Acid / Q000737:chemistry; D014361:Trypsin Inhibitors / Q000737:chemistry; D014599:Uterus / Q000503:physiopathology
D007704:Kininogens; D014361:Trypsin Inhibitors; D010664:Phenylmethylsulfonyl Fluoride; D013776:Tetrathionic Acid; D007611:Aprotinin; D004492:Edetic Acid; D007610:Kallikreins