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Biochim Biophys Acta. 2015 Nov;1853(11 Pt A):2856-69. doi: 10.1016/j.bbamcr.2015.08.007. Epub 2015 Aug 15.

Eps15 homology domain containing protein of Plasmodium falciparum (PfEHD) associates with endocytosis and vesicular trafficking towards neutral lipid storage site.

Author information

1
International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India.
2
International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India; Department of Biosciences, Faculty of Natural Sciences, Jamia Millia Islamia, New Delhi 110 025, India.
3
Department of Biotechnology, All India Institute of Medical Sciences, New Delhi 110 029, India.
4
Department of Biochemistry, Faculty of Science, Jamia Hamdard, New Delhi 110062, India.
5
Department of Biosciences, Faculty of Natural Sciences, Jamia Millia Islamia, New Delhi 110 025, India.
6
International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India. Electronic address: amohd@icgeb.res.in.

Abstract

The human malaria parasite, Plasmodium falciparum, takes up numerous host cytosolic components and exogenous nutrients through endocytosis during the intra-erythrocytic stages. Eps15 homology domain-containing proteins (EHDs) are conserved NTPases, which are implicated in membrane remodeling and regulation of specific endocytic transport steps in eukaryotic cells. In the present study, we have characterized the dynamin-like C-terminal Eps15 homology domain containing protein of P. falciparum (PfEHD). Using a GFP-targeting approach, we studied localization and trafficking of PfEHD in the parasite. The PfEHD-GFP fusion protein was found to be a membrane bound protein that associates with vesicular network in the parasite. Time-lapse microscopy studies showed that these vesicles originate at parasite plasma membrane, migrate through the parasite cytosol and culminate into a large multi-vesicular like structure near the food-vacuole. Co-staining of food vacuole membrane showed that the multi-vesicular structure is juxtaposed but outside the food vacuole. Labeling of parasites with neutral lipid specific dye, Nile Red, showed that this large structure is neutral lipid storage site in the parasites. Proteomic analysis identified endocytosis modulators as PfEHD associated proteins in the parasites. Treatment of parasites with endocytosis inhibitors obstructed the development of PfEHD-labeled vesicles and blocked their targeting to the lipid storage site. Overall, our data suggests that the PfEHD is involved in endocytosis and plays a role in the generation of endocytic vesicles at the parasite plasma membrane, that are subsequently targeted to the neutral lipid generation/storage site localized near the food vacuole.

KEYWORDS:

Endocytosis; Eps15 homology domain (EHD); Food-vacuole; Lipid storage; Malaria; Plasmodium falciparum

PMID:
26284889
DOI:
10.1016/j.bbamcr.2015.08.007
[Indexed for MEDLINE]
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