Amphiphilic behavior of a brain tetrameric acetylcholinesterase form lacking the plasma membrane anchoring domain

Fuentes, María-Elena; Inestrosa, Nibaldo C.

doi:10.1016/0006-8993(92)90919-z

Cited by 5 publications

(3 citation statements)

References 16 publications

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“…As shown previously, monomerization and release of the hydrophobic anchor from tetrameric DS-AChE from human proteolysis (Gennari et al, 1987;Heider and Brodbeck, 1992). Also, proteinase K was shown to release mammalian brain tetrameric AChE from a crude membrane fraction of bovine caudate nucleus, presumably by splitting the hydrophobic anchor (Fuentes and Inestrosa, 1992). We used the same method and monomerized human and bovine brain DS-AChE by limited trypsin treatment.…”

Section: Discussionmentioning

confidence: 82%

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Monoclonal antibodies against brain acetycholinesterases which recognize the subunits bearing the hydrophobic anchor

Liao

Mortensen

Nørgaard‐Pedersen

et al. 1993

European Journal of Biochemistry

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Monoclonal antibodies were raised against amphiphilic detergent-soluble (DS) acetylcholinesterase (AChE) from human brain caudate nucleus. Three mAb, 132-4 (IgG,), 132-5 (IgG,) and 132-6 (IgG,), specific for brain DS-AChE were selected and subcloned. These mAb reacted with native as well as heat-denatured and SDS-denatured DS-AChE, indicating that the epitopes to which mAb bound are continuous determinants. The mAb cross-reacted with DS-AChE from bovine and mouse brain and with brain DS-AChE from river trout (Sufmo rrurra f o m f u r i o ) and lake trout (SuZmo rrurru f o m u Zacusrris). No cross-reaction was detected with the following antigens : salt-soluble (SS) AChE from bovine brain, glycophospholipid-anchored AChE from human and bovine erythrocytes, DS-butyrylcholinesterase and SS-butyrylcholinesterase (BtChE) from the brains of human and bovine, DS-BtChE from chicken and BtChE from human serum. Deglycosylation of brain DSAChE with N-glycosidase F did not abolish the binding of mAb to DS-AChE. After reduction of brain DS-AChE by dithiothreitol, the mAb no longer reacted with the antigen, indicating that a disulfide bridge is important for the epitope. Monomerization of brain DS-AChE by trypsin and limited proteinase K treatment also abolished the binding of mAb to DS-ACE. Sucrose-densitygradient centrifugation showed that mAb reacted only with native tetrameric forms, but not with dimeric and monomeric forms. Western blot, after SDSPAGE under non-reducing conditions, showed that mAb reacted with those subunits carrying the hydrophobic anchor (i.e. tetramers, himers and heavy dimers) but not with those devoid of it (light dimers or monomers). Since mAb 132-4,132-5 and 132-6 recognized DS-ACE from fish up to mammalian brain in the evolutionary tree, it is concluded that the epitope to which these mAb bind, is conserved in nature.

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Section: Discussionmentioning

confidence: 82%

“…Also, proteinase K was shown to release mammalian brain tetrameric AChE from a crude membrane fraction of bovine caudate nucleus, presumably by splitting the hydrophobic anchor (Fuentes and Inestrosa, 1992). We used the same method and monomerized human and bovine brain DSAChE by limited trypsin treatment.…”

Section: Discussionmentioning

confidence: 99%

Monoclonal antibodies against brain acetycholinesterases which recognize the subunits bearing the hydrophobic anchor

Liao

Mortensen

Nørgaard‐Pedersen

et al. 1993

European Journal of Biochemistry

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“…Whether the formation of these particular clusters involves the participation of an aggregating factor (e.g., perhaps with properties similar to those of agrin) remains unknown. In turn, the possible existence of an aggregating factor(s) in the mammalian brain has been explored by studying the behavior of hydrophilic 04 AChE released by proteinase K from crude membrane fractions of bovine caudate nuclei (Fuentes and Inestrosa, 1992). In that study, the solubilized G4 AChE maintained its capacity to form aggregates in detergent-free gradients, a property that was not due to the hydrophobic membraneanchoring domain still linked to the enzyme, nor to the presence of AChE activity trapped in small plasma membrane vesicles.…”

Section: Aggregation/clusteringmentioning

confidence: 99%

Tetrameric (G₄) Acetylcholinesterase: Structure, Localization, and Physiological Regulation

Fernández

Moreno

Inestrosa

1996

Journal of Neurochemistry

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Acetylcholinesterase (AChE), a highly conserved enzyme in the animal kingdom, is distributed throughout a wide range of vertebrate tissues where it is expressed as multiple molecular forms comprising different arrangements of catalytic and structural subunits. The major AChE form in the CNS is an amphiphilic globular tetramer (G 4 AChE) consisting of four identical catalytic subunits attached to cellular membranes by a hydrophobic noncatalytic subunit (P-subunit). This study focuses primarily on current data involving the structure of the G4 AChE P-subunit, the expression and regulation of G4 AChE during development and adulthood, and its role(s) in certain neurological disorders including Alzheimer's disease.

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Does acetylcholinesterase secretion involve an ADAMs-like metallosecretase?

Nalivaeva

Turner

1999

Lett Pept Sci

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SummaryThe ADAMs (A Disintegrin And Metalloprotease-like) family is a large and rapidly expanding group of metalloproteinases with structural similarity. They are normally characterized by the presence of a proteolytic domain and disintegrin and signalling domains. Although 21 ADAMs proteins have been already cloned to date, in most cases their natural substrates are unknown. The best characterized representative of the mammalian ADAMs family is the TNF-~ converting enzyme (TACE). TACE is an integral membrane metalloproteinase that causes the secretion of the active form of TNF-~ from its plasma membrane precursor and thus can be regarded as a membrane protein secretase. Secretion of membrane proteins is a very well documented biological phenomenon and was demonstrated for a diverse range of membrane proteins, two examples being angiotensin converting enzyme (ACE) and Alzheimer's amyloid precursor protein (APP). ACE and APP secretion was shown to possess substantial similarity with the secretion of TNF-~. In the present study, we have attempted to demonstrate that a metalloproteinase might be involved in the shedding of another membrane-bound protein -acetylcholinesterase (ACHE). Secretion of AChE by human neuroblastoma SH-SY5Y cells was found to be inhibited by a selective hydroxamate metalloproteinase inhibitor batimastat (20 #M), and stimulated by carbachol (20/xM), which have previously been shown to regulate the activity of APP c~-secretase in a similar manner. The role of ADAMs proteins in the shedding of molecules from the cell surface is discussed.

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Amphiphilic behavior of a brain tetrameric acetylcholinesterase form lacking the plasma membrane anchoring domain

Cited by 5 publications

References 16 publications

Monoclonal antibodies against brain acetycholinesterases which recognize the subunits bearing the hydrophobic anchor

Monoclonal antibodies against brain acetycholinesterases which recognize the subunits bearing the hydrophobic anchor

Tetrameric (G₄) Acetylcholinesterase: Structure, Localization, and Physiological Regulation

Does acetylcholinesterase secretion involve an ADAMs-like metallosecretase?

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Amphiphilic behavior of a brain tetrameric acetylcholinesterase form lacking the plasma membrane anchoring domain

Cited by 5 publications

References 16 publications

Monoclonal antibodies against brain acetycholinesterases which recognize the subunits bearing the hydrophobic anchor

Monoclonal antibodies against brain acetycholinesterases which recognize the subunits bearing the hydrophobic anchor

Tetrameric (G4) Acetylcholinesterase: Structure, Localization, and Physiological Regulation

Does acetylcholinesterase secretion involve an ADAMs-like metallosecretase?

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Tetrameric (G₄) Acetylcholinesterase: Structure, Localization, and Physiological Regulation