Advertisement

The Relationship between Cholinergic Proteolipid and Proteodetergent in Torpedo Electroplax Membranes

  • E. De Robertis
  • S. de Fiszer Plazas
  • M. C. de Llorente Carlin
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 83)

Abstract

The two methodological approaches used for the separation of the cholinergic receptor from electric tissue: i. e. organic solvents (De Robertis et al., 1967, LaTorre et al., 1970) or detergent extraction (Changeux et al., 1970; Miledi et al., 1971) have led to the isolation of hydrophobic intrinsic proteins from the electroplax membranes. Because of the intimate relationship of these proteins with lipids, in one case, or detergents, in the other, here they will be called “cholinergic proteolipid” and “cholinergic proteodetergent”. The problem which will be considered is that of the identity or non-identity of these proteins; in other words if they are totally different or if they have some similarities. Because both the proteolipid and the proteodetergent have high affinity binding for cholinergic agonists and antagonists of nicotinic type and could be separated from acetylcholinesterase (De Robertis and Fiszer de Plazas, 1970), we were inclined to think that they were similar or at least partially related (Fiszer de Plazas and De Robertis, 1972). Later on, since most of the work on the proteodetergent was based on α-bungarotoxin we studied the binding on this α-toxin to the proteolipid from the electroplax. We found that α-bungarotoxin displaced the site of high affinity for the cholinergic fluorescent probe dansylcholine (De Robertis and Barrantes, 1972). Furthermore, it was possible to demonstrate the binding of labeled α-bungarotoxin to the proteolipid by the use of Sephadex LH 20 chromatography. In experiments done on the electroplax membranes extracted with organic solvents, the membranes lost the binding capacity for α-bungarotoxin, while there was binding to the extracted proteolipid (Fiszer de Plazas and De Robertis, 1972).

Keywords

Affinity Chromatography High Affinity Binding Cholinergic Receptor Cholinergic Agonist Detergent Extraction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barrantes F. J. (1973) Proc. 9th Internat. Congress Biochem. Stockholm, p. 443Google Scholar
  2. Barrantes F. J., Changeux J. P., Lunt G. G. & Sobel A. (1975a) Nature 256, 325–327PubMedCrossRefGoogle Scholar
  3. Barrantes F. J., Ochoa E. L. M., Arbilla S., de Carlin M. C. L. & De Robertis E. (1975b) Biochem, Biophys. Res. Commun. 63, 194–200CrossRefGoogle Scholar
  4. Bonner W. M. & Laskey R. A. (1974) Eur. J. Biochem. 46, 83–88PubMedCrossRefGoogle Scholar
  5. Changeux J. P., Kasai M., Huchet M. & Meunier J. C. (1970) C.R. Acad. Sci. Paris 270, 2864–2867Google Scholar
  6. De Robertis E. (1975) Synaptic Receptors. Isolation and Molecular Biology, Dekker, New YorkGoogle Scholar
  7. De Robertis E. & Barrantes F. J. (1972) Eur. J. Pharmacol. 17, 303–305PubMedCrossRefGoogle Scholar
  8. De Robertis E., Fiszer de Plazas S. & Soto E. F. (1967) Science 158, 928–929PubMedCrossRefGoogle Scholar
  9. De Robertis E. & Fiszer de Plazas S. (1970) Biochem. Biophys. Acta 219, 388–397PubMedCrossRefGoogle Scholar
  10. De Robertis E., Fiszer de Plazas S. & de Carlin M. C. L. (1976) Nature 259, 605–606PubMedCrossRefGoogle Scholar
  11. De Robertis E., Lunt G. S. & LaTorre J. L. (1971) Mol. Pharmacol. 7, 97–103PubMedGoogle Scholar
  12. Eldefrawi M. E. & Eldefrawi A. T. (1973) Arch. Biochem. Biophys. 159, 362–373PubMedCrossRefGoogle Scholar
  13. Fiszer de Plazas S. & De Robertis E. (1972) Biochim. Biophys. Acta 274, 258–265CrossRefGoogle Scholar
  14. Gitler C. (1972) Ann. Rev. Biophys. Bioeng. 1, 51–125CrossRefGoogle Scholar
  15. Karlin A. (1974) Life Sciences 14, 1385–1415PubMedCrossRefGoogle Scholar
  16. Karlin A. & Cowburn D. (1973) Proc. Natl. Acad. Sci. U. S. A. 70, 3636–3640PubMedCrossRefGoogle Scholar
  17. Karlsson E., Helbronn E. & Windlund L. (1972) Febs Letters 28, 107–111PubMedCrossRefGoogle Scholar
  18. Klett R. P., Fulpius B. W., Cooper D., Smith M., Reich E. & Possani L. (1973) J. Biol. Chem. 248, 6841–6853PubMedGoogle Scholar
  19. Laemmli V. K. (1970) Nature 227, 680–685PubMedCrossRefGoogle Scholar
  20. Laskey R. A. & Mills A. D. (1975) Eur. J. Biochem. 56, 335–341PubMedCrossRefGoogle Scholar
  21. LaTorre J. L., Lunt G. S. & De Robertis E. (1970) Proc. Natl. Acad. Sci. U. S. A. 65, 716–720CrossRefGoogle Scholar
  22. Meunier J. C., Sealock R., Olsen R. & Changeux J. P. (1974) Eur. J. Biochem. 45, 371–394PubMedCrossRefGoogle Scholar
  23. Miledi R., Molinoff P. & Potter L. T. (1971) Nature 229, 554–557PubMedCrossRefGoogle Scholar
  24. O’Farrell P. H. (1975) J. Biol. Chem. 250, 4007–4021PubMedGoogle Scholar
  25. Olsen R. W., Meunier J. C. & Changeux J. P. (1972) Febs Letters 28, 96–100PubMedCrossRefGoogle Scholar
  26. Schmidt J. & Raftery M. A. (1972) Biochem. Biophys. Res. Commun. 49, 572–578PubMedCrossRefGoogle Scholar
  27. Studier F. W. (1973) J. Mol. Biol. 79, 237–248PubMedCrossRefGoogle Scholar
  28. Weber G., Borris D. P., De Robertis E., Barrantes J. F., LaTorre J. L. & Llorente de Carlin M. C. (1971) Mol. Pharmacol. 7, 530–537PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1977

Authors and Affiliations

  • E. De Robertis
    • 1
  • S. de Fiszer Plazas
    • 1
  • M. C. de Llorente Carlin
    • 1
  1. 1.Instituto de Biología Celular, Facultad de MedicinaUniversidad de Buenos AiresBuenos AiresArgentina

Personalised recommendations