Biochemical and Biophysical Research Communications, Vol.298, No.4, 559-565, 2002
Cell surface expression of a GPI-anchored form of mouse acetylcholinesterase in Klpmr1 Delta cells of Kluyveromyces lactis
The mouse acetylcholinesterase AChE(H) was expressed in the yeast Kluyveromyces lactis. The AChEH activity was detectable in intact cells whereas it was absent in the culture media. Glucanase treatment and immunoelectron microscopy data indicated that AChEH is anchored to plasma membrane and that the mouse GPI-signaling is compatible with the K. lactis targeting machinery. The AChEH was also expressed in a K lactis strain carrying an inactivated allele of KlPMR1, the gene coding for a P-type Ca2+-ATPase of the Golgi apparatus. This mutant displays changes in protein glycosylation and cell wall structure. The AChEH activity detected in Klpmr1Delta cells was more than twofold higher than that observed in wild-type cells. The combination of ACNE expression and anchoring with the characteristics of Klpmr1Delta strain of K. lactis resulted in yeast cells displaying high ACNE activity. This could be regarded as a novel sensing unit to be employed for detecting ACNE inhibitors as pesticides. (C) 2002 Elsevier Science (USA). All rights reserved.