Protein composition of the atrial muscarinic acetylcholine receptor partially purified by wheat germ agglutinin affinity chromatography

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Abstract

The use of affinity chromatography on wheat germ agglutinin columns to partially purify detergent extracts of muscarinic acetylcholine receptor from porcine atria is described. The procedure results in a 20-fold purification of the protein. The partially purified protein binds [3H]L-QNB (the l isomer of quinuclidinyl benzilate) with an observed association rate constant 2- to 3-fold lower than that found for the detergent extract; however, incubation with column fractions eluted prior to the receptor gives an association rate constant similar to that for detergent extracts. The component responsible for this effect is nondialyzable and protease sensitive, indicating that it may be a protein or high-molecular-weight peptide. Affinity labeling experiments with [3H]propylbenzilylcholine mustard [N. J. M. Birdsall, A. S. V. Burgen, and E. C. Hulme (1979) Brit. J. Pharmacol.66, 337–342] show radioactivity incorporated mainly in a broad peak of apparent molecular weight 75,000 ± 5000.

References (36)

  • A.J. Beld et al.

    Eur. J. Pharmacol

    (1974)
  • O. Hurko

    Arch. Biochem. Biophys

    (1978)
  • M. Gavish et al.

    Biochem. Biophys. Res. Commun

    (1982)
  • C.R. Cremo et al.

    Anal. Biochem

    (1981)
  • J. Wei et al.

    Eur. J. Pharmacol

    (1980)
  • L.B. Rosenberger et al.

    J. Biol. Chem

    (1980)
  • C.P. Berrie et al.

    Biochem. Biophys. Res. Commun

    (1979)
  • O.H. Lowry et al.

    J. Biol. Chem

    (1951)
  • G.L. Peterson

    Anal. Biochem

    (1977)
  • I. Kahane et al.

    Biochim. Biophys. Acta

    (1976)
  • J.B. Galper et al.

    J. Biol. Chem

    (1980)
  • H. Repke et al.

    Pharmazie

    (1980)
  • H. Gorrisen et al.

    Anal. Biochem

    (1981)
  • T. Haga et al.

    Biomed. Res

    (1982)
  • E.C. Hulme et al.

    J. Receptor Res

    (1983)
  • G.L. Peterson et al.

    Prep. Biochem

    (1983)
  • G.S. Herron et al.

    Biochemistry

    (1982)
  • G.S. Herron et al.

    J. Neurochem

    (1983)
  • Cited by (0)

    Supported in part by NIH Grant 23632 from the Heart, Lung and Blood Institute.

    2

    Present address, Laboratory of Radiobiology and Environmental Health, LR102, University of California, San Francisco, Calif. 94143.

    1

    M.I.S. is the recipient of Research Career Development Award HL 00796 from the National Institutes of Health.

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