title: Isolation, characterization and partial purification of a transferable membrane channel (amoebapore) produced by Entamoeba histolytica.
abstract: Entamoeba histolytica kills cells by contact dependent cytolysis. The mechanism underlying this process must be of rapid onset because target cells round up and show marked zeiosis within 15 min of contact. In earlier work, we identified a remarkable ion-channel forming protein which we named amoebapore, that may contribute to the amoeba-induced target cell killing. Within the amoeba it exists as part of a supramolecular aggregate together with other proteins of unknown function. In this work we report the purification of a solubilized form of the amoebapore. Amoebapore was found to exist as an apparent dimer of the previously reported protein whose molecular weight had been determined under denaturing conditions. Two isoforms of this dimer, with pI values of 6.8 and 5.3 present at a ratio of 7 to 1, were identified and purified. Both isoforms demonstrate ion-channel forming activity in planar lipid membranes. These channels show a unit conductance of 5-20 pS and remain open for less than 1 s. Upon lateral aggregation, opening becomes concerted to a greater degree with channel conductance are observed. The isolated particulate form of amoebapore depolarizes cells.
authors: Department of Membrane Research, Weizmann Institute of Science, Rehovot, Israel.
authors: I
authors: I
authors: Rosenberg
authors: D
authors: D
authors: Bach
authors: L M
authors: LM
authors: Loew
authors: C
authors: C
authors: Gitler
mesh_terms: D000818:Animals; D002460:Cell Line; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004748:Entamoeba histolytica / Q000032:analysis* / Q000648:ultrastructure; D005470:Fluorometry; D007473:Ion Channels / Q000032:analysis*; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000032:analysis / Q000302:isolation & purification*; D015800:Protozoan Proteins
chemical_list: D007473:Ion Channels; D008051:Lipid Bilayers; D008565:Membrane Proteins; D015800:Protozoan Proteins; C041670:amoebapore proteins, protozoan