Identification of phosphatidylcholine transfer protein-like in the parasite Entamoeba histolytica

Piña-Vázquez, Carolina; Reyes-López, Magda; Mendoza‐Hernández, Guillermo; Bermúdez‐Cruz, Rosa María; Garza, Mireya de la

doi:10.1016/j.biochi.2014.09.007

Cited by 4 publications

(4 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previously expressed and purified His tag recombinant EhLTP3 (rEhLTP3) and EhLTP1 (rEhLTP1) were used to study their abilities to bind with lipid molecules as per previously described protocol with slight modifications [57]. Briefly, each Membrane Lipid Strip (P-6002, Echelon Bioscience Inc.) where the following lipids were spotted: triglyceride (GT), diacylglycerol (DAG), phosphatidic Acid (PA), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylglycerol (PG), cardiolipin (CL), phosphatidylinositol (PtdIns), PtdIns 4-phosphate [PI(4)P] (PtdIns phosphate called phosphoinositide, designated as PI), PtdIns 4,5-bisphosphate [PtdIns(4,5)P 2 ], PtdIns 3,4,5-trisphosphate [PtdIns (3,4,5)P 3 ], cholesterol, sphingomyelin (SM), 3-sulfogalactosylceramide or sulfatide (SGC), and Blue Blank; was blocked with 3% BSA in PBS for 2h at RT prior to incubation with either rEhLTP3 or rEhLTP1 (1mg/mL) in PBS with 3% BSA for 16 h at 4˚C.…”

Section: Protein-lipid Overlay Assaymentioning

confidence: 99%

Two StAR-related lipid transfer proteins play specific roles in endocytosis, exocytosis, and motility in the parasitic protist Entamoeba histolytica

2021

View full text Add to dashboard Cite

Lipid transfer proteins (LTPs) are the key contributor of organelle-specific lipid distribution and cellular lipid homeostasis. Here, we report a novel implication of LTPs in phagocytosis, trogocytosis, pinocytosis, biosynthetic secretion, recycling of pinosomes, and motility of the parasitic protist E. histolytica, the etiological agent of human amoebiasis. We show that two StAR-related lipid transfer (START) domain-containing LTPs (named as EhLTP1 and 3) are involved in these biological pathways in an LTP-specific manner. Our findings provide novel implications of LTPs, which are relevant to the elucidation of pathophysiology of the diseases caused by parasitic protists.

show abstract

Section: Protein-lipid Overlay Assaymentioning

confidence: 99%

Two StAR-related lipid transfer proteins play specific roles in endocytosis, exocytosis, and motility in the parasitic protist Entamoeba histolytica

2021

View full text Add to dashboard Cite

show abstract

“…E. histolytica possesses full capacity of de novo biosynthesis of phospholipids including phosphatidylcholine and phosphatidylethanolamine [52] and the ability to scavenge components from the environment under in vitro cultivation conditions [53]. Furthermore, Entamoeba possesses a list of lipid binding proteins in their genome [54,55]; one of such lipid transfer proteins, EhPCTP-L, was shown to bind phosphatidylserine and phosphatidic acid and localizes to the cell surface [56]. The catalytic P4-ATPase subunit of the phospholipid flipase responsible for miltefosine transport should be identified to better understand the molecular mechanisms of phospholipid transport on the plasma membrane and its involvement in drug resistant and pathogenesis in Entamoeba .…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of the Entamoeba Histolytica Rab8a Binding Protein: A Cdc50 Homolog

Hanadate

Saito‐Nakano

Nakada‐Tsukui

et al. 2018

IJMS

View full text Add to dashboard Cite

Membrane traffic plays a pivotal role in virulence in the enteric protozoan parasite Entamoeba histolytica. EhRab8A small GTPase is a key regulator of membrane traffic at the endoplasmic reticulum (ER) of this protist and is involved in the transport of plasma membrane proteins. Here we identified the binding proteins of EhRab8A. The Cdc50 homolog, a non-catalytic subunit of lipid flippase, was identified as an EhRab8A binding protein candidate by affinity coimmunoprecipitation. Binding of EhRab8A to EhCdc50 was also confirmed by reciprocal immunoprecipitation and blue-native polyacrylamide gel electrophoresis, the latter of which revealed an 87 kDa complex. Indirect immunofluorescence imaging with and without Triton X100 showed that endogenous EhCdc50 localized on the surface in the absence of permeabilizing agent but was observed on the intracellular structures and overlapped with the ER marker Bip when Triton X100 was used. Overexpression of N-terminal HA-tagged EhCdc50 impaired its translocation to the plasma membrane and caused its accumulation in the ER. As reported previously in other organisms, overexpression and accumulation of Cdc50 in the ER likely inhibited surface transport and function of the plasma membrane lipid flippase P4-ATPase. Interestingly, HA-EhCdc50-expressing trophozoites gained resistance to miltefosine, which is consistent with the prediction that HA-EhCdc50 overexpression caused its accumulation in the ER and mislocalization of the unidentified lipid flippase. Similarly, EhRab8A gene silenced trophozoites showed increased resistance to miltefosine, supporting EhRab8A-dependent transport of EhCdc50. This study demonstrated for the first time that EhRab8A mediates the transport of EhCdc50 and lipid flippase P4-ATPase from the ER to the plasma membrane.

show abstract

“…However, non-vesicular lipid transport machinery and the function of LTPs remained largely unexplored in E. histolytica . There are two previous reports where the lipid trafficking in E. histolytica was described (Pina-Vázquez et al, 2014 ; Bolaños et al, 2016 ). Piña- Vázquez et al identified a START domain containing protein in E. histolytica (EHI_110720, also present in our list of E. histolytica LTP homologs) and named it as E. histolytica phosphatidylcholine transfer protein-like (EhPCTP-L).…”

Section: Biological Significance Of Lipid Transfer In E Himentioning

confidence: 93%

“…They identified EhPCTP-L by virtue of interaction with anti-chicken embryo caveolin-1 monoclonal antibody, which indicates their potential role in caveola-mediated endocytosis. EhPCTP-L mainly binds to anionic phospholipids phosphatidylserine (PS) and PA, and is localized to the PM and the cytosol (Pina-Vázquez et al, 2014 ). However, the essential biochemical characteristics as LTPs, i.e., lipid transport activity, was not reported in this study.…”

Section: Biological Significance Of Lipid Transfer In E Himentioning

confidence: 99%

Non-vesicular Lipid Transport Machinery in Entamoeba histolytica

Das

Nozaki

2018

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Eukaryotic cells are organized into separate membrane-bound compartments that have specialized biochemical signature and function. Maintenance and regulation of distinct identity of each compartment is governed by the uneven distribution and intra-cellular movement of two essential biomolecules, lipids, and proteins. Non-vesicular lipid transport mediated by lipid transfer proteins plays a pivotal role in intra-cellular lipid trafficking and homeostasis whereas vesicular transport plays a central role in protein trafficking. Comparative study of lipid transport machinery in protist helps to better understand the pathogenesis and parasitism, and provides insight into eukaryotic evolution. Amebiasis, which is caused by Entamoeba histolytica, is one of the major enteric infections in humans, resulting in 40–100 thousand deaths annually. This protist has undergone remarkable alterations in the content and function of its sub-cellular compartments as well represented by its unique diversification of mitochondrion-related organelle, mitosome. We conducted domain-based search on AmoebaDB coupled with bioinformatics analyses and identified 22 potential lipid transfer protein homologs in E. histolytica, which are grouped into several sub-classes. Such in silico analyses have demonstrated the existence of well-organized lipid transport machinery in this parasite. We summarized and discussed the conservation and unique features of the whole repertoire of lipid transport proteins in E. histolytica.

show abstract

Identification of phosphatidylcholine transfer protein-like in the parasite Entamoeba histolytica

Cited by 4 publications

References 53 publications

Two StAR-related lipid transfer proteins play specific roles in endocytosis, exocytosis, and motility in the parasitic protist Entamoeba histolytica

Two StAR-related lipid transfer proteins play specific roles in endocytosis, exocytosis, and motility in the parasitic protist Entamoeba histolytica

Identification and Characterization of the Entamoeba Histolytica Rab8a Binding Protein: A Cdc50 Homolog

Non-vesicular Lipid Transport Machinery in Entamoeba histolytica

Contact Info

Product

Resources

About