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. 1990 Dec;9(12):3885–3888. doi: 10.1002/j.1460-2075.1990.tb07607.x

Anionic subsites of the acetylcholinesterase from Torpedo californica: affinity labelling with the cationic reagent N,N-dimethyl-2-phenyl-aziridinium.

C Weise 1, H J Kreienkamp 1, R Raba 1, A Pedak 1, A Aaviksaar 1, F Hucho 1
PMCID: PMC552156  PMID: 2249655

Abstract

Several peptides of acetylcholinesterase of Torpedo californica labelled with the alkylating reagent [3H]N,N-dimethyl-2-phenyl-aziridinium (DPA) were localized within the primary structure. One peptide had the sequence KPQELIDVE (positions 270-278); the incorporation of DPA into this peptide could be specifically suppressed by propidium, which suggests that it is part of the peripheral anionic site. The incorporation of DPA into two other peptides was insensitive to propidium but could be prevented by edrophonium; the sequence of one of the peptides assumed to be part of the anionic site in the catalytic centre was found to be DLFR (positions 217-220). Decamethonium efficiently blocked alkylation by DPA in all three investigated peptides.

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Selected References

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  1. Belleau B., Tani H. A novel irreversible inhibitor of acetylcholinesterase specifically directed at the anionic binding site: structure-activity relationships. Mol Pharmacol. 1966 Sep;2(5):411–422. [PubMed] [Google Scholar]
  2. Brady L., Brzozowski A. M., Derewenda Z. S., Dodson E., Dodson G., Tolley S., Turkenburg J. P., Christiansen L., Huge-Jensen B., Norskov L. A serine protease triad forms the catalytic centre of a triacylglycerol lipase. Nature. 1990 Feb 22;343(6260):767–770. doi: 10.1038/343767a0. [DOI] [PubMed] [Google Scholar]
  3. Changeux J. P. Responses of acetylcholinesterase from Torpedo marmorata to salts and curarizing drugs. Mol Pharmacol. 1966 Sep;2(5):369–392. [PubMed] [Google Scholar]
  4. Chothia C., Leuzinger W. Acetylcholinesterase: the structure of crystals of a globular form from electric eel. J Mol Biol. 1975 Sep 5;97(1):55–60. doi: 10.1016/s0022-2836(75)80021-0. [DOI] [PubMed] [Google Scholar]
  5. Dudai Y., Silman I., Shinitzky M., Blumberg S. Purification by affinity chromatography of the molecular forms of acetylcholinesterase present in fresh electric-organ tissue of electric eel. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2400–2403. doi: 10.1073/pnas.69.9.2400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. ELLMAN G. L., COURTNEY K. D., ANDRES V., Jr, FEATHER-STONE R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961 Jul;7:88–95. doi: 10.1016/0006-2952(61)90145-9. [DOI] [PubMed] [Google Scholar]
  7. Futerman A. H., Low M. G., Silman I. A hydrophobic dimer of acetylcholinesterase from Torpedo californica electric organ is solubilized by phosphatidylinositol-specific phospholipase C. Neurosci Lett. 1983 Sep 19;40(1):85–89. doi: 10.1016/0304-3940(83)90097-6. [DOI] [PubMed] [Google Scholar]
  8. Krupka R. M. Hydrolysis of neutral substrates by acetylcholinesterase. Biochemistry. 1966 Jun;5(6):1983–1988. doi: 10.1021/bi00870a028. [DOI] [PubMed] [Google Scholar]
  9. Kupke T., Lechner M., Kaim G., Götz F. Improved purification and biochemical properties of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis. Eur J Biochem. 1989 Oct 20;185(1):151–155. doi: 10.1111/j.1432-1033.1989.tb15096.x. [DOI] [PubMed] [Google Scholar]
  10. Lockridge O., Bartels C. F., Vaughan T. A., Wong C. K., Norton S. E., Johnson L. L. Complete amino acid sequence of human serum cholinesterase. J Biol Chem. 1987 Jan 15;262(2):549–557. [PubMed] [Google Scholar]
  11. MacPhee-Quigley K., Taylor P., Taylor S. Primary structures of the catalytic subunits from two molecular forms of acetylcholinesterase. A comparison of NH2-terminal and active center sequences. J Biol Chem. 1985 Oct 5;260(22):12185–12189. [PubMed] [Google Scholar]
  12. NACHMANSOHN D., WILSON I. B. The enzymic hydrolysis and synthesis of acetylcholine. Adv Enzymol Relat Subj Biochem. 1951;12:259–339. doi: 10.1002/9780470122570.ch5. [DOI] [PubMed] [Google Scholar]
  13. Nolte H. J., Rosenberry T. L., Neumann E. Effective charge on acetylcholinesterase active sites determined from the ionic strength dependence of association rate constants with cationic ligands. Biochemistry. 1980 Aug 5;19(16):3705–3711. doi: 10.1021/bi00557a011. [DOI] [PubMed] [Google Scholar]
  14. Purdie J. E. The properties of acetylcholinesterase modified by interaction with the alkylating agent N,N-dimethyl-2-phenylaziridinium ion. Biochim Biophys Acta. 1969 Jul 8;185(1):122–133. doi: 10.1016/0005-2744(69)90288-5. [DOI] [PubMed] [Google Scholar]
  15. Rosenberry T. L. Acetylcholinesterase. Adv Enzymol Relat Areas Mol Biol. 1975;43:103–218. doi: 10.1002/9780470122884.ch3. [DOI] [PubMed] [Google Scholar]
  16. Schrag J. D., Schmid M. F., Morgan D. G., Phillips G. N., Jr, Chiu W., Tang L. Crystallization and preliminary X-ray diffraction analysis of 11 S acetylcholinesterase. J Biol Chem. 1988 Jul 15;263(20):9795–9800. [PubMed] [Google Scholar]
  17. Sussman J. L., Harel M., Frolow F., Varon L., Toker L., Futerman A. H., Silman I. Purification and crystallization of a dimeric form of acetylcholinesterase from Torpedo californica subsequent to solubilization with phosphatidylinositol-specific phospholipase C. J Mol Biol. 1988 Oct 5;203(3):821–823. doi: 10.1016/0022-2836(88)90213-6. [DOI] [PubMed] [Google Scholar]
  18. Taylor P., Lappi S. Interaction of fluorescence probes with acetylcholinesterase. The site and specificity of propidium binding. Biochemistry. 1975 May 6;14(9):1989–1997. doi: 10.1021/bi00680a029. [DOI] [PubMed] [Google Scholar]
  19. Weise C., Kreienkamp H. J., Raba R., Aaviksaar A., Hucho F. The active site and partial sequence of cobra venom acetylcholinesterase. J Protein Chem. 1990 Feb;9(1):53–57. doi: 10.1007/BF01024984. [DOI] [PubMed] [Google Scholar]
  20. Winkler F. K., D'Arcy A., Hunziker W. Structure of human pancreatic lipase. Nature. 1990 Feb 22;343(6260):771–774. doi: 10.1038/343771a0. [DOI] [PubMed] [Google Scholar]

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