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Molecular comparison of α2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelet

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Summary

We have previously described a simple two-step purification technique to isolate α2-adrenergic receptors from the rat adrenocortical carcinoma (Jaiswal, R. K. and Sharma, R. K. (1985) Biochem. Biophys. Res. Commun. 130, 58–64). Utilizing this technique we have now achieved ∼ 77 000-fold purification to apparent homogeneity of α2-adrenergic receptors from human platelets. We have compared the biochemical characteristics of these receptors with those from the rat, which were purified ∼ 40000-fold to homogeneity.

The [125I] receptor proteins from two sources showed: (a) a single radioactive band with a Mr of 64000 as evidenced by one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE); and (b) a single symmetrical peak with a pl of 4.2 by isoelectric focusing polyacrylamide gel electrophoresis. Both proteins showed typical α2-adrenergic binding characteristics with specific binding activities of 13.85 nmol/mg and 14.17 nmol/mg protein. These values are close to the theoretical binding activity of 15.6 nmol/mg protein for 1 mol of the ligand binding 1 mol of the receptor protein. These results attest to the purity of the receptors, to its Mr of 64000, and to its acidic nature. However, the peptide maps of the radioiodinated α2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelets reveal some distinct differences which may relate to the differences in the pharmacological specificities between rodent and nonrodent α2-adrenergic receptors.

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Abbreviations

PAC:

p-aminoclonidine

PMSF:

Phenylmethyl-sulfonylfluoride

DTT:

Dithiothreitol

HPLC:

High Performance Liquid Chromatography

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Authors and Affiliations

  1. Department of Biochemistry, University of Tennessee, 38163, Memphis, TN, USA

    Rama Kant Jaiswall & Rameshwar K. Sharma

  2. Cold Spring Harbor Laboratories, 11724, Cold Spring Harbor, NY, USA

    Daniel R. Marshak

Authors
  1. Rama Kant Jaiswall
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  2. Daniel R. Marshak
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  3. Rameshwar K. Sharma
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Jaiswall, R.K., Marshak, D.R. & Sharma, R.K. Molecular comparison of α2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelet. Mol Cell Biochem 86, 41–53 (1989). https://doi.org/10.1007/BF00231688

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  • DOI: https://doi.org/10.1007/BF00231688

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