Anticoagulant repertoire of the hookworm Ancylostoma caninum.
Hookworms are hematophagous nematodes that infect a wide range of mammalian hosts, including humans. There has been speculation for nearly a century as to the identity of the anticoagulant substances) used by these organisms to subvert host hemostasis. Using molecular cloning, we describe a family of potent small protein (75-84 amino acids) anticoagulants from the hookworm Ancylostoma caninum termed AcAP (A. caninum anticoagulant protein). Two recombinant AcAP members (AcAP5 and AcAP6) directly inhibited the catalytic activity of blood coagulation factor Xa (fXa), while a third form (AcAPc2) predominantly inhibited the catalytic activity of a complex composed of blood coagulation factor VIIa and tissue factor (fVIIa/TF). The inhibition of fVIIa/TF was by a unique mechanism that required the initial formation of a binary complex of the inhibitor with fXa at a site on the enzyme that is distinct from the catalytic center (exo-site). The sequence of AcAPc2 as well as the utilization of an exo-site on fXa distinguishes this inhibitor from the mammalian anticoagulant TFPI (tissue factor pathway inhibitor), which is functionally equivalent with respect to fXa-dependent inhibition of fIIa/TF. The relative sequence positions of the reactive site residues determined for AcAP5 with the homologous regions in AcAP6 and AcAPc2 as well as the pattern of 10 cysteine residues present in each of the inhibitors suggest that the AcAPs are distantly related to the family of small protein serine protease inhibitors found in the nonhematophagous nematode Ascaris lumbricoides var. suum.
Stassens
P
P
Corvas International Inc., San Diego, CA 92121, USA.
Bergum
P W
PW
Gansemans
Y
Y
Jespers
L
L
Laroche
Y
Y
Huang
S
S
Maki
S
S
Messens
J
J
Lauwereys
M
M
Cappello
M
M
Hotez
P J
PJ
Lasters
I
I
Vlasuk
G P
GP
D000595:Amino Acid Sequence; D000722:Ancylostoma / Q000201:enzymology* / Q000235:genetics; D000818:Animals; D001665:Binding Sites; D001777:Blood Coagulation; D001779:Blood Coagulation Factors / Q000037:antagonists & inhibitors*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015801:Helminth Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D015842:Serine Proteinase Inhibitors / Q000235:genetics*; D013925:Thromboplastin / Q000378:metabolism
D001779:Blood Coagulation Factors; D018076:DNA, Complementary; D015801:Helminth Proteins; D015842:Serine Proteinase Inhibitors; D013925:Thromboplastin