Existence of two membrane-bound acetylcholinesterases in the honey bee head.
Two acetylcholinesterase (EC 3.1.1.7) membrane forms AChE(m1) and AChE(m2), have been characterised in the honey bee head. They can be differentiated by their ionic properties: AChE(m1) is eluted at 220 mM NaCl whereas AChE(m2) is eluted at 350 mM NaCl in anion exchange chromatography. They also present different thermal stabilities. Previous processing such as sedimentation, phase separation, and extraction procedures do not affect the presence of the two forms. Unlike AChE(m1), AChE(m2) presents reversible chromatographic elution properties, with a shift between 350 to 220 mM NaCl, depending on detergent conditions. Purification by affinity chromatography does not abolish the shift of the AChE(m2) elution. The similar chromatographic behaviour of soluble AChE strongly suggests that the occurrence of the two membrane forms is not due to the membrane anchor. The two forms have similar sensitivities to eserine and BW284C51. They exhibit similar electrophoretic mobilities and present molecular masses of 66 kDa in SDS-PAGE and a sensitivity to phosphatidylinositol-specific phospholipase C in non-denaturing conditions, thus revealing the presence of a glycosyl-phosphatidylinositol anchor. We assume that bee AChE occurs in two distinct conformational states whose AChE(m2) apparent state is reversibly modulated by the Triton X-100 detergent into AChE(m1).
Badiou
Alexandra
A
INRA, Laboratoire de Toxicologie Environnementale, UMR INRA-UAPV 406 Ecologie des Invertébrés, Avignon, France. alexandra.badiou@avignon.inra.fr
Brunet
Jean-Luc
JL
Belzunces
Luc P
LP
D000110:Acetylcholinesterase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001516:Bees / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006257:Head; D019476:Insect Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017830:Octoxynol / Q000737:chemistry; D054801:Phosphoinositide Phospholipase C / Q000378:metabolism
D019476:Insect Proteins; D008565:Membrane Proteins; D017830:Octoxynol; D000110:Acetylcholinesterase; D054801:Phosphoinositide Phospholipase C