Metalloproteases of infective Ancylostoma hookworm larvae and their possible functions in tissue invasion and ecdysis.
To infect their hosts, hookworm larvae must exsheath and migrate through connective tissue. A modified in vitro skin chamber was used to show that the human hookworm Ancylostoma duodenale and the zoonotic canine hookworm Ancylostoma caninum penetrate epidermis, basement membrane, and dermis in similar ways. These similarities in tissue invasion properties reflect the observed biochemical similarities in parasite protease composition. The larvae of both species contain protease activity that is inhibited by o-phenanthroline; this identifies the proteases as metalloproteases. The enzyme activities exhibit an alkaline pH optimum between pH 9 and 10. During modified sodium dodecyl sulfate-polyacrylamide gel electrophoresis in which a protein substrate (either casein or gelatin) was used, the protease activities resolved into a major band at an Mr of 68,000 and a minor band at an Mr of 38,000. Proteases were released by living A. caninum larvae in vitro and degraded purified and radiolabeled casein to smaller peptides. Motile hookworm larvae were also incubated with purified and radiolabeled connective tissue macromolecules in vitro. Both Ancylostoma species degraded human fibronectin to a 60,000-Mr polypeptide intermediate, but could not degrade solubilized bovine elastin or human laminin. In contrast, the obligate skin-penetrating nematode Strongyloides stercoralis degraded all three substrates. This biochemical difference may explain some observed differences in invasiveness.
Hotez
P
P
Yale MacArthur Center for Molecular Parasitology, Yale University School of Medicine, New Haven, Connecticut 06510.
Haggerty
J
J
Hawdon
J
J
Milstone
L
L
Gamble
H R
HR
Schad
G
G
Richards
F
F
D000722:Ancylostoma / Q000201:enzymology* / Q000502:physiology; D000818:Animals; D003238:Connective Tissue / Q000469:parasitology; D004285:Dogs; D006801:Humans; D007814:Larva / Q000201:enzymology; D008666:Metalloendopeptidases / Q000032:analysis* / Q000502:physiology; D008970:Molecular Weight; D012867:Skin / Q000469:parasitology
D008666:Metalloendopeptidases