title: Identification of differentially expressed water-insoluble proteins in the encystment process of Colpoda cucullus by two-dimensional electrophoresis and LC-MS/MS analysis.
abstract: In the encystment process of the ciliate protist Colpoda cucullus, we observed that the cell total protein abundance was reduced at 12 h-1 d after the onset of encystment induction subsequent to the reduction in mRNA abundance. We analyzed the alteration of the expression levels of water-insoluble proteins by two-dimensional polyacrylamide gel electrophoresis using polyoxyethylene (20) sorbitan monooleate (Tween-80), and we identified proteins whose expression levels were altered in the encystment process by a liquid chromatography tandem mass spectrometry analysis. The expression level of a 60-kDa protein (p60; heat shock protein 60) was temporarily enhanced and that of a 55-kDa protein (p55; actin) and a 49-kDa protein (p49; actin) was enhanced in the Colpoda encystment process. In mature cysts, the expression level of p55 and p49 tended to be reduced, whereas the expression level of a 50-kDa protein (p50d; α-tubulin), a 25-kDa protein (p25; α-tubulin) and a 52-kDa protein (p52c; β-tubulin) was enhanced.
authors.affiliation: Department of Biological Science, Faculty of Science, Kochi University, Kochi, 780-8520, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Sogame
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Kojima
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Takeshita
authors.forename: Eiji
authors.initials: E
authors.lastname: Kinoshita
authors.forename: Tatsuomi
authors.initials: T
authors.lastname: Matsuoka
mesh_terms: D000199:Actins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D000818:Animals; D018834:Chaperonin 60 / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D002853:Chromatography, Liquid; D016798:Ciliophora / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development*; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D033761:Spores, Protozoan / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development*; D053719:Tandem Mass Spectrometry
chemical_list: D000199:Actins; D018834:Chaperonin 60; D010446:Peptide Fragments; D015800:Protozoan Proteins; D012333:RNA, Messenger; C095402:peptide 277, heat shock protein 60
keywords: ATP synthase; Actin; cellular differentiation; cryptobiosis; cyst formation; heat shock protein; tubulin