A. Madi et al., BIOCHEMICAL-CHARACTERIZATION AND LOCALIZATION OF TRANSGLUTAMINASE IN WILD-TYPE AND CELL-DEATH MUTANTS OF THE NEMATODE CAENORHABDITIS-ELEGANS, European journal of biochemistry, 253(3), 1998, pp. 583-590
Transglutaminase activity was characterized in extracts of the nematod
e Caenorhabditis elegans using a microtiter plate method, and found to
be Ca2+-dependent, optimal at pH 8.0, and to be inhibited by EGTA, am
monia, iodoacetamide and GTP. Monoclonal and polyclonal antibodies rai
sed against human tissue transglutaminase also inhibited the activity
and detected a 61-kDa protein from the worm lysate. Constitutive expre
ssion of the enzyme in the wild-type intestinal cells was revealed by
immunohistochemistry. Potential protein substrates for the enzyme were
found in worm lysates using a biotin-labelled amine substrate. There
is a basal level of protein-bound epsilon(gamma-glutamyl)lysine cross-
links, characteristic of transglutaminase activity, formed in situ in
adult wild-type animals. Developmental studies have revealed that the
enzyme activity is highest in adult animals, and relatively higher in
L1 larvae than in other larval stages. As compared to wild types, lowe
r transglutaminase activity has been measured in lysates of ced-3, ced
-4 and ced-9 mutants. Cross-link levels were also low in ced-4 and ced
-9 mutants. By contrast, the crosslink content was high in several pha
gocytosis mutants. The highest concentration was found in the ced-5; c
ed-7 double phagocytosis mutants which carry an extra number of dead c
ells during their lifespan. In accordance with this finding, several t
ransglutaminase-immunopositive cells were found in both the embryos an
d in the head of these double phagocytosis mutants. The results sugges
t that a transglutaminase is involved in, or related to, the death pro
gram of cells in C. elegans and the expression and crosslinking activi
ty of the enzyme may be perturbed in some ced mutants.