Two distinct proteins are associated with tetrameric acetylcholinesterase on the cell surface

Anselme L. Perrier, Xavier Cousin, Nicola Boschetti, Robert Haas, Jean Marc Chatel, Suzanne Bon, William L. Roberts, Samuel R. Pickett, Jean Massoulié, Terrone L. Rosenberry, Eric Krejci

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

In mammalian brain, acetylcholinesterase (ACHE) exists mostly as a tetramer of 70-kDa catalytic subunits that are linked through disulfide bonds to a hydrophobic subunit P of approximately 20 kDa. To characterize P, we reduced the disulfide bonds in purified bovine brain AChE and sequenced tryptic fragments from bands in the 20-kDa region. We obtained sequences belonging to at least two distinct proteins: the P protein and another protein that was not disulfide-linked to catalytic subunits. Both proteins were recognized in Western blots by antisera raised against specific peptides. We cloned cDNA encoding the second protein in a cDNA library from bovine substantia nigra and obtained rat and human homologs. We call this protein mCutA because of its homology to a bacterial protein (CutA). We could not demonstrate a direct interaction between mCutA and AChE in vitro in transfected cells. However, in a mouse neuroblastoma cell line that produced membrane-bound AChE as an amphiphilic tetramer, the expression of mCutA antisense mRNA eliminated cell surface AChE and decreased the level of amphiphilic tetramer in cell extracts. mCutA therefore appears necessary for the localization of AChE at the cell surface; it may be part of a multicomponent complex that anchors AChE in membranes, together with the hydrophobic P protein.

Original languageEnglish (US)
Pages (from-to)34260-34265
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number44
StatePublished - Nov 3 2000

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Acetylcholinesterase
Disulfides
Proteins
Brain
Catalytic Domain
Membranes
Bacterial Proteins
Substantia Nigra
Cell Extracts
Anchors
Neuroblastoma
Gene Library
Rats
Immune Sera
Complementary DNA
Western Blotting
Cells
Cell Line
Messenger RNA
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Perrier, A. L., Cousin, X., Boschetti, N., Haas, R., Chatel, J. M., Bon, S., ... Krejci, E. (2000). Two distinct proteins are associated with tetrameric acetylcholinesterase on the cell surface. Journal of Biological Chemistry, 275(44), 34260-34265.

Two distinct proteins are associated with tetrameric acetylcholinesterase on the cell surface. / Perrier, Anselme L.; Cousin, Xavier; Boschetti, Nicola; Haas, Robert; Chatel, Jean Marc; Bon, Suzanne; Roberts, William L.; Pickett, Samuel R.; Massoulié, Jean; Rosenberry, Terrone L.; Krejci, Eric.

In: Journal of Biological Chemistry, Vol. 275, No. 44, 03.11.2000, p. 34260-34265.

Research output: Contribution to journalArticle

Perrier, AL, Cousin, X, Boschetti, N, Haas, R, Chatel, JM, Bon, S, Roberts, WL, Pickett, SR, Massoulié, J, Rosenberry, TL & Krejci, E 2000, 'Two distinct proteins are associated with tetrameric acetylcholinesterase on the cell surface', Journal of Biological Chemistry, vol. 275, no. 44, pp. 34260-34265.
Perrier AL, Cousin X, Boschetti N, Haas R, Chatel JM, Bon S et al. Two distinct proteins are associated with tetrameric acetylcholinesterase on the cell surface. Journal of Biological Chemistry. 2000 Nov 3;275(44):34260-34265.
Perrier, Anselme L. ; Cousin, Xavier ; Boschetti, Nicola ; Haas, Robert ; Chatel, Jean Marc ; Bon, Suzanne ; Roberts, William L. ; Pickett, Samuel R. ; Massoulié, Jean ; Rosenberry, Terrone L. ; Krejci, Eric. / Two distinct proteins are associated with tetrameric acetylcholinesterase on the cell surface. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 44. pp. 34260-34265.
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