Identification of a novel transglutaminase from the filarial parasite Brugia malayi and its role in growth and development.
Recently, we reported the presence of a putative transglutaminase in adult female worms of Brugia malayi [1]. The enzyme activity was shown to be essential for in utero growth and development of microfilariae. Here, we demonstrate that adult worms of B. malayi have a large amount of epsilon-(gamma-glutamyl)lysine isopeptide bonds, a product of physiologically active transglutaminase. A 25-kDa immunoreactive band detected in female worm extracts by a monospecific monoclonal antibody (CUB 7401) against guinea pig liver transglutaminase was associated with the enzymatic activity. Unlike the mammalian enzyme, the parasite enzyme did not require Ca2+ for its catalytic activity. Furthermore, in utero developing embryos, especially during early stages of development, contained very high amounts of this enzyme. Adult female worms contained several proteins that could serve as suitable substrates for the enzyme. Inhibition of the enzyme activity by an enzyme-specific pseudosubstrate, monodansylcadaverine, led to a time- and dose-dependent inhibition of microfilariae production and release by gravid female worms. The inhibition of microfilariae production was due to the inhibition of transglutaminase-catalyzed crosslinking of parasite proteins that in turn seemed to be essential for in utero growth and development of the embryos. The results suggest that transglutaminase-catalyzed reactions may play an important role during early development of embryos to mature microfilariae inside the adult female worms of filarial parasites.
Department of Medical Oncology, University of Texas, M.D. Anderson Cancer Center, Houston 77030.
K
K
Mehta
U R
UR
Rao
A C
AC
Vickery
L
L
Fesus
D000818:Animals; D002009:Brugia / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D015801:Helminth Proteins / Q000378:metabolism; D008297:Male; D008842:Microfilariae / Q000187:drug effects / Q000378:metabolism; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000378:metabolism*
D015801:Helminth Proteins; D011503:Transglutaminases; C008542:monodansylcadaverine; D002103:Cadaverine