title: Entamoeba histolytica: analysis of the trophozoite proteome by two-dimensional polyacrylamide gel electrophoresis.
abstract: The Entamoeba histolytica genome project carried out at TIGR and the Sanger Institute has produced a near-complete set of deduced open reading frame sequences. These data provide strong support for the identification of signals from proteomic analyses such as two-dimensional electrophoresis by protein sequencing and/or mass spectrometric methods. To carry out an initial investigation of the E. histolytica proteome, appropriate sample preparation and silver staining protocols were adapted. After preparation of protein extracts from E. histolytica HM-1:IMSS trophozoites, solubilized proteins were separated in the first dimension in IPG (immobilized pH gradient) strips depending on their pI and subsequently in the second dimension according to their molecular weight by SDS-polyacrylamide gel electrophoresis. Of the more than 1500 protein spots visualized, several landmark spots were isolated from the gels and identified by either tryptic cleavage and subsequent MALDI-TOF mass spectrometry or by protein sequencing.
authors.affiliation: Department of Specific Prophylaxis and Tropical Medicine, Center for Physiology and Pathophysiology, Kinderspitalg. 15, A-1095 Vienna, Austria.
authors.forename: David
authors.initials: D
authors.lastname: Leitsch
authors.forename: Christian
authors.initials: C
authors.lastname: Radauer
authors.forename: Katharina
authors.initials: K
authors.lastname: Paschinger
authors.forename: Iain B H
authors.initials: IB
authors.lastname: Wilson
authors.forename: Heimo
authors.initials: H
authors.lastname: Breiteneder
authors.forename: Otto
authors.initials: O
authors.lastname: Scheiner
authors.forename: Michael
authors.initials: M
authors.lastname: Duchêne
mesh_terms: D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D004748:Entamoeba histolytica / Q000737:chemistry / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry; D020539:Sequence Analysis, Protein; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D020543:Proteome; D015800:Protozoan Proteins