Abstract
In the collagen-tailed forms of cholinesterases, each subunit of a specific triple helical collagen, ColQ, may be attached through a proline-rich domain (PRAD) situated in its N-terminal noncollagenous region, to tetramers of acetylcholinesterase (AChE) or butyrylcholinesterase (BChE). This heteromeric assembly ensures the functional anchoring of AChE in extracellulare matrices, for example, at the neuromuscular junction. In this study, we analyzed the influence of deletions in the noncollagenous C-terminal region of ColQ on its capacity to form a triple helix. We show that an 80-residue segment located downstream of the collagenous regions contains the trimerization domain, that it can form trimers without the collagenous regions, and that a pair of cysteines located at the N-boundary of this domain facilitates oligomerization, although it is not absolutely required. We further show that AChE subunits can associate with nonhelical collagen ColQ monomers, forming ColQ-associated tetramers (G4-Q), which are secreted or are anchored at the cell surface when the C-terminal domain of ColQ is replaced by a GPI-addition signal.
Similar content being viewed by others
REFERENCES
Massoulié, J., Pezzementi, L., Bon, S., Krejci, E., and Vallette, F. M. 1993. Molecular and cellular biology of cholinesterases. Prog. Neurobiol. 41:31–91.
Massoulié, J., Anselmet, A., Bon, S., Krejci, E., Legay, C., Morel, N., and Simon, S. 1998. Acetylcholinesterase: C-terminal domains, molecular forms and functional localization. J. Physiol. (Paris) 92:183–190.
Massoulié, J. 2002. The origin of the molecular diversity and functional anchoring of cholinesterases. NeuroSignals 11:130–143.
Bon, S., Rosenberry, T. L., and Massoulié, J. 1991. Amphiphilic, glycophosphatidylinositol-specific phospholipase C (PI-PLC)-insensitive monomers and dimers of acetylcholinesterase. Cell. Mol. Neurobiol. 11:157–172.
Bon, S. and Massoulié, J. 1997. Quaternary associations of acetylcholinesterase: I. Oligomeric associations of T subunits with and without the amino-terminal domain of the collagen tail. J. Biol. Chem. 272:3007–3015.
Krejci, E., Coussen, F., Duval, N., Chatel, J. M., Legay, C., Puype, M., Vandekerckhove, J., Cartaud, J., Bon, S., and Massoulié, J. 1991. Primary structure of a collagenic tail peptide of Torpedo acetylcholinesterase: Co-expression with catalytic subunit induces the production of collagen-tailed forms in transfected cells. EMBO J. 10:1285–1293.
Krejci, E., Thomine, S., Boschetti, N., Legay, C., Sketelj, J., and Massoulié, J. 1997. The mammalian gene of acetylcholinesterase-associated collagen. J. Biol. Chem. 272:22840–22847.
Feng, G., Krejci, E., Molgo, J., Cunningham, J. M., Massoulié, J., and Sanes, J. R. 1999. Genetic analysis of collagen Q: Roles in acetylcholinesterase and butyrylcholinesterase assembly and in synaptic structure and function. J. Cell Biol. 144:1349–1360.
Perrier, A. L., Massoulié, J., and Krejci, E. 2002. PRiMA, the membrane anchor of acetylcholinesterase in brain. Neuron 33:275–285.
Hall, Z. W. 1973. Multiple forms of acetylcholinesterase and their distribution in endplate and non-endplate regions of rat diaphragm muscle. J. Neurol. 4:343–361.
Gennari, K., Brunner, J., and Brodbeck, U. 1987. Tetrameric detergent-soluble acetylcholinesterase from human caudate nucleus: Subunit composition and number of active sites. J. Neurochem. 49:12–18.
Inestrosa, N. C., Roberts, W. L., Marshall, T. L., and Rosenberry, T. L. 1987. Acetylcholinesterase from bovine caudate nucleus is attached to membranes by a novel subunit distinct from those of acetylcholinesterases in other tissues. J. Biol. Chem. 262:4441–4444.
Bon, S., Coussen, F., and Massoulié, J. 1997. Quaternary associations of acetylcholinesterase: II. The polyproline attachment domain of the collagen tail. J. Biol. Chem. 272:3016–3021.
Simon, S., Krejci, E., and Massoulié, J. 1998. A four-to-one association between peptide motifs: Four C-terminal domains from cholinesterase assemble with one proline-rich attachment domain (PRAD) in the secretory pathway. EMBO J. 17:6178–6187.
Engel, J. and Prockop, D. J. 1991. The zipper-like folding of collagen triple helices and the effects of mutations that disrupt the zipper. Annu. Rev. Biophys. Biophys. Chem. 20:137–152.
McLaughlin, S. H. and Bulleid, N. J. 1998. Molecular recognition in procollagen chain assembly. Matrix Biol. 16:369–377.
Duval, N., Krejci, E., Grassi, J., Coussen, F., Massoulié, J., and Bon, S. 1992. Molecular architecture of acetylcholinesterase collagen-tailed forms: Construction of a glycolipid-tailed tetramer. EMBO J. 11:3255–3261.
Legay, C., Bon, S., Vernier, P., Coussen, F., and Massoulié, J. 1993. Cloning and expression of a rat acetylcholinesterase subunit: Generation of multiple molecular forms and complementarity with a Torpedo collagenic subunit. J. Neurochem. 60:337–346.
Mizushima, S. and Nagata, S. 1990. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 18:5322.
Selden, R. F., Howie, K. B., Rowe, M. E., Goodman, H. M., and Moore, D. D. 1986. Human growth hormone as a reporter gene in regulation studies employing transient gene expression. Mol. Cell Biol. 6:3173–3179.
Bon, S. and Massoulié, J. 1978. Collagenase sensitivity and aggregation properties of Electrophorus acetylcholinesterase. Eur. J. Biochem. 89:89–94.
Ellman, G. L., Courtney, K. D., Andres, V., and Featherstone, R. M. 1961. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7:88–95.
Bon, S., Toutant, J. P., Méflah, K., and Massoulié, J. 1988. Amphiphilic and nonamphiphilic forms of Torpedo cholinesterases: II. Electrophoretic variants and phosphatidylinositol phospholipase C-sensitive and-insensitive forms. J. Neurochem. 51:786–794.
Karnovsky, M. J. and Roots, L. 1964. A direct-coloring thiocholine method for cholinesterases. J. Histochem. Cytochem. 12:219–222.
Rost, B. 1996. PHD: Predicting one-dimensional protein structure by profile-based neural networks. Methods Enzymol. 266:525–539.
Bon, S., Cartaud, J., and Massoulié, J. 1978. Dumbbell-shaped associations of tailed Electrophorus acetylcholinesterase molecules. Mol. Biol. Rep. 4:61–63.
Beck, K., Boswell, B. A., Ridgway, C. C., and Bachinger, H. P. 1996. Triple helix formation of procollagen type I can occur at the rough endoplasmic reticulum membrane. J. Biol. Chem. 271:21566–21573.
Bulleid, N. J., Dalley, J. A., and Lees, J. F. 1997. The C-propeptide domain of procollagen can be replaced with a transmembrane domain without affecting trimer formation or collagen triple helix folding during biosynthesis. EMBO J. 16:6694–6701.
Sikorav, J. L., Duval, N., Anselmet, A., Bon, S., Krejci, E., Legay, C., Osterlund, M., Reimund, B., and Massoulié, J. 1988. Complex alternative splicing of acetylcholinesterase transcripts in Torpedo electric organ: Primary structure of the precursor of the glycolipid-anchored dimeric form. EMBO J. 7:2983–2993.
Duval, N., Massoulié, J., and Bon, S. 1992. H and T subunits of acetylcholinesterase from Torpedo, expressed in COS cells, generate all types of globular forms. J. Cell Biol. 118:641–653.
Coussen, F., Ayon, A., Le Goff, A., Leroy, J., Massoulié, J., and Bon, S. 2001. Addition of a glycophosphatidylinositol to acetylcholinesterase: Processing, degradation, and secretion. J. Biol. Chem. 276:27881–27892.
Kristensen, T., Oxvig, C., Sand, O., Moller, N. P., and Sottrup-Jensen, L. 1994. Amino acid sequence of human pregnancy-associated plasma protein-A derived from cloned cDNA. Biochemistry 33:1592–1598.
Corpet, F., Gouzy, J., and Kahn, D. 1998. The ProDom database of protein domain families. Nucleic Acids Res. 26:323–326.
Lawrence, J. B., Oxvig, C., Overgaard, M. T., Sottrup-Jensen, L., Gleich, G. J., Hays, L. G., Yates, J. R., 3rd, and Conover, C. A. 1999. The insulin-like growth factor (IGF)-dependent IGF binding protein-4 protease secreted by human fibroblasts is pregnancy-associated plasma protein-A. Proc. Natl. Acad. Sci. USA 96:3149–3153.
Overgaard, M. T., Haaning, J., Boldt, H. B., Olsen, I. M., Laursen, L. S., Christiansen, M., Gleich, G. J., Sottrup-Jensen, L., Conover, C. A., and Oxvig, C. 2000. Expression of recombinant human pregnancy-associated plasma protein-A and identification of the proform of eosinophil major basic protein as its physiological inhibitor. J. Biol. Chem. 275:31128–31133.
Mechling, D. E., Gambee, J. E., Morris, N. P., Sakai, L. Y., Keene, D. R., Mayne, R., and Bächinger, H. P. 1996. Type IX collagen NC1 domain peptides can trimerize in vitro without forming a triple helix. J. Biol. Chem. 271:13781-13785.
Brass, A., Kadler, K. E., Thomas, J. T., Grant, M. E., and Boot-Handford, R. P. 1992. The fibrillar collagens, collagen VIII, collagen X and the C1q complement proteins share a similar domain in their C-terminal non-collagenous regions. FEBS Lett. 303:126–128.
Hoppe, H. J., Barlow, P. N., and Reid, K. B. 1994. A parallel three stranded alpha-helical bundle at the nucleation site of collagen triple-helix formation. FEBS Lett. 344:191–195.
Moradi-Ameli, M., Deleage, G., Geourjon, C., and van der Rest, M. 1994. Common topology within a non-collagenous domain of several different collagen types. Matrix Biol. 14:233–239.
Lesage, A., Penin, F., Geourjon, C., Marion, D., and van der Rest, M. 1996. Trimeric assembly and three-dimensional structure model of the FACIT collagen COL1-NC1 junction from CD and NMR analysis. Biochemistry. 35:9647–9660.
Kao, W. W., Prockop, D. J., and Berg, R. A. 1979. Kinetics for the secretion of nonhelical procollagen by freshly isolated tendon cells. J. Biol. Chem. 254:2234–2243.
Thakker-Varia, S., Anderson, D. W., Kuivaniemi, H., Tromp, G., Shin, H. G. van der Rest, M., Glorieux, F. H., Ala-Kokko, L., and Stolle, C. A. 1995. Aberrant splicing of the type III procollagen mRNA leads to intracellular degradation of the protein in a patient with Ehlers-Danlos type IV. Hum. Mutat. 6:116–125.
Lamandé, S. R., Chessler, S. D., Golub, S. B., Byers, P. H., Chan, D., Cole, W. G., Sillence, D. O., and Bateman, J. F. 1995. Endoplasmic reticulum-mediated quality control of type I collagen production by cells from osteogenesis imperfecta patients with mutations in the pro alpha 1 (I) chain carboxyl-terminal propeptide which impair subunit assembly. J. Biol. Chem. 270:8642–8649.
Walmsley, A. R., Batten, M. R., Lad, U., and Bulleid, N. J. 1999. Intracellular retention of procollagen within the endoplasmic reticulum is mediated by prolyl 4-hydroxylase. J. Biol. Chem. 274:14884–14892.
Bon, S., Cartaud, J., and Massoulié, J. 1978. The dependence of acetylcholinesterase aggregation at low ionic strength upon a polyanionic component. Eur. J. Biochem. 85:1–14.
Deprez, P. and Inestrosa, N. C. 1995. Two heparin-binding domains are present on the collagenic tail of asymmetric acetylcholinesterase. J. Biol. Chem. 270:11043–11046.
Deprez, P., Doss-Pepe, E., Brodsky, B., and Inestrosa, N. C. 2000. Interaction of the collagen-like tail of asymmetric acetylcholinesterase with heparin depends on triple-helical conformation, sequence and stability. Biochem. J. 350:283–290.
Kuivaniemi, H., Tromp, G., and Prockop, D. J. 1997. Mutations in fibrillar collagens (types I, II, III, and XI), fibril-associated collagen (type IX), and network-forming collagen (type X) cause a spectrum of diseases of bone, cartilage, and blood vessels. Hum. Mutat. 9:300–315.
Ohno, K., Brengman, J., Tsujino, A., and Engel, A. G. 1998. Human endplate acetylcholinesterase deficiency caused by mutations in the collagen-like (ColQ) of the asymmetric enzyme. Proc. Natl. Acad. Sci. USA 95:9654–9659.
Ohno, K., Brengman, J. M., Felice, K. J., Cornblath, D. R., and Engel, A. G. 1999. Congenital endplate acetylcholinesterase deficiency caused by a nonsense mutation and an A→G splicedonor-site mutation at position +3 of the collagenlike-tail-subunit gene (COLQ): How does G at position +3 result in aberrant splicing? Am. J. Hum. Genet. 65:635–644.
Ohno, K., Engel, A. G., Brengman, J. M., Shen, X. M., Heidenreich, F., Vincent, A., Milone, M., Tan, E., Demirci, M., Walsh, P., Nakano, S., and Akiguchi, I. 2000. The spectrum of mutations causing endplate acetylcholinesterase deficiency. Ann. Neurol. 47:162–170.
Shapira, Y. A., Sadeh, M. E., Bergtraum, M. P., Tsujino, A., Ohno, K., Shen, X. M., Brengman, J., Edwardson, S., Matoth, I., and Engel, A. G. 2002. Three novel COLQ mutations and variation of phenotypic expressivity due to G240X. Neurology 58:603–609.
Ohno, K. and Engel, A. G. 2002. Congenital myasthenic syndromes: Genetic defects of the neuromuscular junction. Curr. Neurol. Neurosci. Rep. 2:78–88.
Rossi, S. G. and Rotundo, R. L. 1996. Transient interactions between collagen-tailed acetylcholinesterase and sulfated proteoglycans prior to immobilization in the extracellular matrix. J. Biol. Chem. 271:1979–1987.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bon, S., Ayon, A., Leroy, J. et al. Trimerization Domain of the Collagen Tail of Acetylcholinesterase. Neurochem Res 28, 523–535 (2003). https://doi.org/10.1023/A:1022821306722
Issue Date:
DOI: https://doi.org/10.1023/A:1022821306722