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Primary structure of an agonist binding subunit of the nicotinic acetylcholine receptor from bovine adrenal chromaffin cells

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Abstract

Activation by acetylcholine of a nicotinic acetylcholine receptor on the membrane of bovine chromaffin cells leads to membrane depolarization and to the subsequent triggering of catecholamine secretion. It is evident that acetylcholine receptors play a central role in the initial phase of the secretion process and, therefore, an extensive characterization of their molecular components and properties is of fundamental interest. With this intention, we have screened bovine adrenal medullary cDNA libraries with a probe coding for a fragment of the rat muscle acetylcholine receptor α subunit. Several cDNA clones were isolated. The longest cDNA had an open reading frame encoding a 495-amino acid protein with a molecular weight of 56,911. The deduced primary structure contains features that indicate that the encoded protein is an α or acetylcholine binding subunit, and, in fact, it manifests significant sequence similarity to previously cloned α subunits. Sequence identity is particularly high with the α3 subunit, which is expressed in the rat pheochromocytoma PC12 cell line and in several brain areas, and consequently, it is considered a component of a neuronal acetylcholine receptor. Accordingly, the present results suggest that the agonist binding subunit of the nicotinic acetylcholine receptor from bovine chromaffin cells is an α3-type subunit, corroborating previous immunological and pharmacological evidence for the presence of a neuronal nicotinic receptor in chromaffin cells.

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Abbreviations

nAChR:

nicotinic acetylcholine receptor

SDS:

sodium dodecyl sulfate

SSC:

0.15 M NaCl and 0.015 M sodium citrate

kb:

kilobases

bp:

base pairs

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Authors and Affiliations

  1. Departmento de Neuroquímica and Instituto de Neurociencias, Universidad de Alicante, Ap. 374, E-03080, Alicante, (Spain)

    Manuel Criado & Angel Navarro

  2. Max-Planck-Institut für Biophys. Chemie, Göttingen, (Fed. Rep. of Germany)

    Manuel Criado & Leonor Alamo

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  1. Manuel Criado
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  2. Leonor Alamo
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Criado, M., Alamo, L. & Navarro, A. Primary structure of an agonist binding subunit of the nicotinic acetylcholine receptor from bovine adrenal chromaffin cells. Neurochem Res 17, 281–287 (1992). https://doi.org/10.1007/BF00966671

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