INVESTIGADORES
TOUZ Maria Carolina
congresos y reuniones científicas
Título:
Clathrin-associated protein in Giardia lamblia.
Autor/es:
FELIZIANI C; ZAMPONI N; MIRAS S; ROPOLO AS; TOUZ MC
Lugar:
San Luis
Reunión:
Congreso; SAIB; 2011
Resumen:
The presence of clathrin-associated proteins (CLASP) has not yet
been reported in the protozoan G. lamblia. However, by searching in
the GDB, we found a protein that contains an ENTH domain
characteristic of the CLASP epsin. The analysis of the participation
of the ENTH protein (ENTHp) in the trafficking towards
lysosomallike peripheral vacuoles (PVs) was initiated by cloning
and expression of the HA-fusion protein and its mutants in GiardiaG. lamblia. However, by searching in
the GDB, we found a protein that contains an ENTH domain
characteristic of the CLASP epsin. The analysis of the participation
of the ENTH protein (ENTHp) in the trafficking towards
lysosomallike peripheral vacuoles (PVs) was initiated by cloning
and expression of the HA-fusion protein and its mutants in GiardiaGiardia
trophozoites. Using IFA and confocal microscopy we showed that
ENTHp localize mainly in the cytosol and around the nuclei,
partially colocalizing with BIP and AP-2. Additionally, we
developed monoclonal antibodies directed against clathrin heavy
chain (CHC) to disclose the interaction and function of these
particular proteins. Clathrin positive polyclonal Ab (pAb)
production was tested in wild-type trophozoites observing that the
pAb recognized a protein localized in the PVs and a band of 203 kDa
corresponding to the CHC. Using this pAb we observed the
colocalization of ENTHp and CHC in the PVs and also validate the
interaction between both proteins. Antibody-secreting hybridomas
for CLH will allowed a better characterization of clathrin and also
corroborate whether the ENTHp is a CLASP acting as a dual epsin-
EpsinR protein. These results will greatly increase our
understanding of unique aspects of lysosomal trafficking in
Giardia..