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Specificity of the Rat Vesicular Acetylcholine Transporter

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Abstract

The protein kinase A–deficient PC12 cell line PC12A123.7 lacks both choline acetyltransferase and the vesicular acetylcholine transporter. This cell line has been used to establish a stably transfected cell line expressing recombinant rat vesicular acetylcholine transporter that is appropriately trafficked to small synaptic vesicles. Acetylcholine is transported by the rat vesicular acetylcholine transporter at a maximal rate of 1.45 nmol acetylcholine/min/mg protein and exhibits a Km for transport of 2.5 mM. The transporter binds vesamicol with a Kd of 7.5 nM. The ability of structural analogs of acetylcholine to inhibit both acetylcholine uptake and vesamicol binding was measured. The results demonstrate that like Torpedo vesicular acetylcholine transporter, the mammalian transporter can bind a diverse group of acetylcholine analogs.

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REFERENCES

  1. Parsons, S. M., Prior, C., and Marshall, I. G. 1993. Acetylcholine transport, storage, and release. Int. Rev. Neurobiol. 35:279–389.

    PubMed  Google Scholar 

  2. Usdin, T. B., Eiden, L. E., Bonner, T. I., and Erickson, J. D. 1995. Molecular biology of the vesicular ACh transporter. Trends Neurosci. 18:218–224.

    PubMed  Google Scholar 

  3. Bahr, B. A. and Parsons, S. M. 1986. Acetylcholine transport and drug inhibition kinetics in Torpedo synaptic vesicles. J. Neurochem. 46:1214–1218.

    PubMed  Google Scholar 

  4. Bahr, B. A. and Parsons, S. M. 1986. Demonstration of a receptor in Torpedo synaptic vesicles for the acetylcholine storage blocker L-trans-2-(4-phenyl[3,4-3H]-piperidino) cyclohexanol. Proc. Natl. Acad. Sci. USA 83:2267–2270.

    PubMed  Google Scholar 

  5. Bahr, B., Clarkson, E. D., Rogers, G. A., Noremberg, K., and Parsons, S. M. 1992. A kinetic and allosteric model for the acetylcholine transporter-vesamicol receptor in synaptic vesicles. Biochemistry 31:5752–5762.

    PubMed  Google Scholar 

  6. Diebler, M.-F. and Morot Gaudry-Talarmain, Y. 1989. AH5183 and cetiedil: Two potent inhibitors of acetylcholine uptake into isolated synaptic vesicles from Torpedo marmorata. J. Neurochem. 52:813–821.

    PubMed  Google Scholar 

  7. Kim, M. H., Lu, M., Lim, E. J., Chai, Y. G., and Hersh, L. B. 1999. Mutational analysis of aspartate residues in the transmembrane regions and cytoplasmic loops of rat vesicular acetylcholine transporter. J. Biol. Chem. 274:673–680.

    PubMed  Google Scholar 

  8. Kim, M. H., Lu, M., Kelly, M., Hersh, L. B. 2000. Mutational analysis of basic residues in the rat vesicular acetylcholine transporter: Identification of a transmembrane ion pair and evidence that histidine is not involved in proton translocation. J. Biol. Chem. 275:6175–6180.

    PubMed  Google Scholar 

  9. Rogers, A. and Parsons, S. M. 1989. Inhibition of acetylcholine storage by acetylcholine analogs in vitro. Mol. Pharmacol. 36:333–341.

    PubMed  Google Scholar 

  10. Clarkson, E. D., Rogers, A., and Parsons, S. M. 1992. Binding and active transport of large analogues of acetylcholine by cholinergic synaptic vesicles in vitro. J. Neurochem. 59:695–700.

    PubMed  Google Scholar 

  11. Ginty, D. D., Glowacka, D., Defranco, C., and Wagner, J. A. 1991. Nerve growth factor-induced neuronal differentiation after dominant repression of both type I and type II cAMP-dependent protein kinase activities. J. Biol. Chem. 266:15325–15333.

    PubMed  Google Scholar 

  12. Inoue, H., Li, Y-P., Wagner, J. A., and Hersh, L. B. 1995. Expression of the choline acetyltransferase gene depends on protein kinase A activity. J. Neurochem. 64:985–990.

    PubMed  Google Scholar 

  13. Shimojo, M., Wu, D., and Hersh, L. B. 1998. The cholinergic gene locus (choline acetyltransferase and acetylcholine transporter genes) is coordinately regulated by protein kinase A II. J. Neurochem. 71:1118–1126.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, 40536–0084

    Myung-Hee Kim & Mei Lu

  2. Department of Chemistry, University of California at Santa Barbara, Santa Barbara, California, 93106

    Gary Rogers & Stanley Parsons

  3. Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, 40536–0084

    Louis B. Hersh

Authors
  1. Myung-Hee Kim
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  2. Mei Lu
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  3. Gary Rogers
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  4. Stanley Parsons
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  5. Louis B. Hersh
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Correspondence to Louis B. Hersh.

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Kim, MH., Lu, M., Rogers, G. et al. Specificity of the Rat Vesicular Acetylcholine Transporter. Neurochem Res 28, 473–476 (2003). https://doi.org/10.1023/A:1022804903088

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  • DOI: https://doi.org/10.1023/A:1022804903088

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