title: Cell surface expression of a GPI-anchored form of mouse acetylcholinesterase in Klpmr1Delta cells of Kluyveromyces lactis.
abstract: The mouse acetylcholinesterase AChE(H) was expressed in the yeast Kluyveromyces lactis. The AChE(H) activity was detectable in intact cells whereas it was absent in the culture media. Glucanase treatment and immunoelectron microscopy data indicated that AChE(H) is anchored to plasma membrane and that the mouse GPI-signaling is compatible with the K. lactis targeting machinery. The AChE(H) was also expressed in a K. lactis strain carrying an inactivated allele of KlPMR1, the gene coding for a P-type Ca(2+)-ATPase of the Golgi apparatus. This mutant displays changes in protein glycosylation and cell wall structure. The AChE(H) activity detected in Klpmr1Delta cells was more than twofold higher than that observed in wild-type cells. The combination of AChE expression and anchoring with the characteristics of Klpmr1Delta strain of K. lactis resulted in yeast cells displaying high AChE activity. This could be regarded as a novel sensing unit to be employed for detecting AChE inhibitors as pesticides.
authors.affiliation: Department of Developmental and Cell Biology, University of Rome La Sapienza, Piazza Aldo More 5, 00185 Rome, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Uccelletti
authors.forename: Antonella
authors.initials: A
authors.lastname: De Jaco
authors.forename: Francesca
authors.initials: F
authors.lastname: Farina
authors.forename: Patrizia
authors.initials: P
authors.lastname: Mancini
authors.forename: Gabriella
authors.initials: G
authors.lastname: Augusti-Tocco
authors.forename: Stefano
authors.initials: S
authors.lastname: Biagioni
authors.forename: Claudio
authors.initials: C
authors.lastname: Palleschi
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D007716:Kluyveromyces / Q000235:genetics*; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D017261:Glycosylphosphatidylinositols; D011994:Recombinant Proteins; D000110:Acetylcholinesterase