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

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

lysosomal–like peripheral vacuoles (PVs) was initiated by cloning

and expression of the HA-fusion protein and its mutants in Giardia

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