title: A novel saposin-like protein of Entamoeba histolytica with membrane-fusogenic activity.
abstract: Amoebapores, the pore-forming proteins of Entamoeba histolytica, are major pathogenicity factors of the parasite. Upon a comprehensive survey in the recently completed genome data sets for the protozoon, we identified in addition to the three amoebapore genes, 16 genes which are constitutively expressed and code for structurally similar proteins, all belonging to the family of saposin-like proteins. Here, we recombinantly expressed in bacteria a defined single entity of this expansive amoebic protein family, namely SAPLIP 3. The protein consists of the saposin-like domain only, comparable to amoebapores, and we characterized its interactions with membranes using different assays. In contrast to amoebapores, SAPLIP 3 neither forms pores in liposomes nor permeabilizes bacterial membranes. However, SAPLIP 3 induces leaky fusion of lipid vesicles as evidenced by fluorescence microscopic analysis and by using a fusion assay that monitors the dequenching of a lipophilic dye. The membrane-fusogenic activity of SAPLIP 3 which is dependent on the presence of negatively charged lipids and on acidic pH resembles in combination with the negative surface charge of the protein characteristics of human saposin C. Beside its function as a cofactor of sphingolipid hydrolysing enzymes, the human protein is considered to be involved in the reorganization of lysosomal compartments due to its fusogenic activity. We hypothesize that in the amoeba, SAPLIP 3 fulfils a similar function in the multifarious endo- and exocytotic transport processes.
authors.affiliation: Research Center for Infectious Diseases, University of Wuerzburg, Roentgenring 11, D-97070 Wuerzburg, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Winkelmann
authors.forename: Matthias
authors.initials: M
authors.lastname: Leippe
authors.forename: Heike
authors.initials: H
authors.lastname: Bruhn
mesh_terms: D000818:Animals; D004705:Endocytosis; D004748:Entamoeba histolytica / Q000378:metabolism / Q000502:physiology*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007473:Ion Channels / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008561:Membrane Fusion; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D049231:Saposins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D007473:Ion Channels; D008081:Liposomes; D015800:Protozoan Proteins; D049231:Saposins; C041670:amoebapore proteins, protozoan