H and T subunits of acetylcholinesterase from Torpedo, expressed in COS cells, generate all types of globular forms

doi:10.1083/jcb.118.3.641

This Article

	Full Text (PDF, 2747K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Duval, N.
	Articles by Bon, S.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Duval, N.
	Articles by Bon, S.


Social Bookmarking

	What's this?

ARTICLES

H and T subunits of acetylcholinesterase from Torpedo, expressed in COS cells, generate all types of globular forms

N Duval, J Massoulie and S Bon
Laboratoire de Neurobiologie, Centre National de la Recherche Scientifique UA 295, Paris, France.

We analyzed the production of Torpedo marmorata acetylcholinesterase (AChE) in transfected COS cells. We report that the presence of an aspartic acid at position 397, homologous to that observed in other cholinesterases and related enzymes (Krejci, E., N. Duval, A. Chatonnet, P. Vincens, and J. Massoulie. 1991. Proc. Natl. Acad. Sci. USA. 88:6647-6651), is necessary for catalytic activity. The presence of an asparagine in the previously reported cDNA sequence (Sikorav, J.L., E. Krejci, and J. Massoulie. 1987. EMBO (Eur. Mol. Biol. Organ.) J. 6:1865-1873) was most likely due to a cloning error (codon AAC instead of GAC). We expressed the T and H subunits of Torpedo AChE, which differ in their COOH-terminal region and correspond respectively to the collagen-tailed asymmetric forms and to glycophosphatidylinositol-anchored dimers of Torpedo electric organs, as well as a truncated T subunit (T delta), lacking most of the COOH- terminal peptide. The transfected cells synthesized similar amounts of AChE immunoreactive protein at 37 degrees and 27 degrees C. However AChE activity was only produced at 27 degrees C and, even at this temperature, only a small proportion of the protein was active. We analyzed the molecular forms of active AChE produced at 27 degrees C. The H polypeptides generated glycophosphatidylinositol-anchored dimers, resembling the corresponding natural AChE form. The cells also released non-amphiphilic dimers G2na. The T polypeptides generated a series of active forms which are not produced in Torpedo electric organs: G1a, G2a, G4a, and G4na cellular forms and G2a and G4na secreted forms. The amphiphilic forms appeared to correspond to type II forms (Bon, S., J. P. Toutant, K. Meflah, and J. Massoulie. 1988. J. Neurochem. 51:776- 785; Bon, S., J. P. Toutant, K. Meflah, and J. Massoulie. 1988. J. Neurochem. 51:786-794), which are abundant in the nervous tissue and muscles of higher vertebrates (Bon, S., T. L. Rosenberry, and J. Massoulie. 1991. Cell. Mol. Neurobiol. 11:157-172). The H and T catalytic subunits are thus sufficient to account for all types of known AChE forms. The truncated T delta subunit yielded only non- amphiphilic monomers, demonstrating the importance of the T COOH- terminal peptide in the formation of oligomers, and in the hydrophobic character of type II forms.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Camp, S., De Jaco, A., Zhang, L., Marquez, M., De La Torre, B., Taylor, P. (2008). Acetylcholinesterase Expression in Muscle Is Specifically Controlled by a Promoter-Selective Enhancesome in the First Intron. J. Neurosci. 28: 2459-2470 [Abstract] [Full Text]
De Jaco, A., Comoletti, D., Kovarik, Z., Gaietta, G., Radic, Z., Lockridge, O., Ellisman, M. H., Taylor, P. (2006). A Mutation Linked with Autism Reveals a Common Mechanism of Endoplasmic Reticulum Retention for the {alpha},beta-Hydrolase Fold Protein Family. J. Biol. Chem. 281: 9667-9676 [Abstract] [Full Text]
Falasca, C., Perrier, N., Massoulie, J., Bon, S. (2005). Determinants of the t Peptide Involved in Folding, Degradation, and Secretion of Acetylcholinesterase. J. Biol. Chem. 280: 878-886 [Abstract] [Full Text]
Birikh, K. R., Sklan, E. H., Shoham, S., Soreq, H. (2003). Interaction of "readthrough" acetylcholinesterase with RACK1 and PKCbeta II correlates with intensified fear-induced conflict behavior. Proc. Natl. Acad. Sci. USA 100: 283-288 [Abstract] [Full Text]
Morel, N., Massoulie, J. (2000). Comparative Expression of Homologous Proteins. A NOVEL MODE OF TRANSCRIPTIONAL REGULATION BY THE CODING SEQUENCE FOLDING COMPATIBILITY OF CHIMERAS. J. Biol. Chem. 275: 7304-7312 [Abstract] [Full Text]
Legay, C., Mankal, F. A., Massoulie, J., Jasmin, B. J. (1999). Stability and Secretion of Acetylcholinesterase Forms in Skeletal Muscle Cells. J. Neurosci. 19: 8252-8259 [Abstract] [Full Text]
Simon, S., Le Goff, A., Frobert, Y., Grassi, J., Massoulie, J. (1999). The Binding Sites of Inhibitory Monoclonal Antibodies on Acetylcholinesterase. IDENTIFICATION OF A NOVEL REGULATORY SITE AT THE PUTATIVE "BACK DOOR". J. Biol. Chem. 274: 27740-27746 [Abstract] [Full Text]
Morel, N., Bon, S., Greenblatt, H. M., Van Belle, D., Wodak, S. J., Sussman, J. L., Massoulié, J., Silman, I. (1999). Effect of Mutations within the Peripheral Anionic Site on the Stability of Acetylcholinesterase. Mol. Pharmacol. 55: 982-992 [Abstract] [Full Text]
Feng, G., Krejci, E., Molgo, J., Cunningham, J. M., Massoulie, J., Sanes, J. R. (1999). Genetic Analysis of Collagen Q: Roles in Acetylcholinesterase and Butyrylcholinesterase Assembly and in Synaptic Structure and Function. JCB 144: 1349-1360 [Abstract] [Full Text]
Bourne, Y., Taylor, P., Bougis, P. E., Marchot, P. (1999). Crystal Structure of Mouse Acetylcholinesterase. A PERIPHERAL SITE-OCCLUDING LOOP IN A TETRAMERIC ASSEMBLY. J. Biol. Chem. 274: 2963-2970 [Abstract] [Full Text]
Luo, Z. D., Camp, S., Mutero, A., Taylor, P. (1998). Splicing of 5' Introns Dictates Alternative Splice Selection of Acetylcholinesterase Pre-mRNA and Specific Expression during Myogenesis. J. Biol. Chem. 273: 28486-28495 [Abstract] [Full Text]
Cousin, X., Bon, S., Massoulie, J., Bon, C. (1998). Identification of a Novel Type of Alternatively Spliced Exon from the Acetylcholinesterase Gene of Bungarus fasciatus. MOLECULAR FORMS OF ACETYLCHOLINESTERASE IN THE SNAKE LIVER AND MUSCLE. J. Biol. Chem. 273: 9812-9820 [Abstract] [Full Text]
Sternfeld, M., Ming, G.-l., Song, H.-j., Sela, K., Timberg, R., Poo, M.-m., Soreq, H. (1998). Acetylcholinesterase Enhances Neurite Growth and Synapse Development through Alternative Contributions of Its Hydrolytic Capacity, Core Protein, and Variable C Termini. J. Neurosci. 18: 1240-1249 [Abstract] [Full Text]
Simon, S., Massoulie, J. (1997). Cloning and Expression of Acetylcholinesterase from Electrophorus. SPLICING PATTERN OF THE 3' EXONS IN VIVO AND IN TRANSFECTED MAMMALIAN CELLS. J. Biol. Chem. 272: 33045-33055 [Abstract] [Full Text]
Shin, I., Kreimer, D., Silman, I., Weiner, L. (1997). Membrane-promoted unfolding of acetylcholinesterase: A possible mechanism for insertion into the lipid�bilayer. Proc. Natl. Acad. Sci. USA 94: 2848-2852 [Abstract] [Full Text]
Cousin, X., Bon, S., Duval, N., Massoulie, J., Bon, C. (1996). Cloning and Expression of Acetylcholinesterase from Bungarus fasciatus Venom. A NEW TYPE OF COOH-TERMINAL DOMAIN; INVOLVEMENT OF A POSITIVELY CHARGED RESIDUE IN THE PERIPHERAL SITE. J. Biol. Chem. 271: 15099-15108 [Abstract] [Full Text]
Perrier, A. L., Cousin, X., Boschetti, N., Haas, R., Chatel, J.-M., Bon, S., Roberts, W. L., Pickett, S. R., Massoulie, J., Rosenberry, T. L., Krejci, E. (2000). Two Distinct Proteins Are Associated with Tetrameric Acetylcholinesterase on the Cell Surface. J. Biol. Chem. 275: 34260-34265 [Abstract] [Full Text]