A simple, fast, one-step method for the purification of the skeletal-muscle ryanodine receptor

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22 Scopus citations

Abstract

In this paper we describe a simple, fast, one-step method for the purification of the skeletal-muscle ryanodine receptor. The ryanodine receptor from CHAPS-solubilized junctional sarcoplasmic-reticulum membranes was adsorbed to a spermine-agarose column and eluted by 2 mM-spermine. The purified receptor, consisting predominantly of a 450 kDa polypeptide on SDS/PAGE, binds [3H]ryanodine with a specific activity of ~ 300 pmol/mg of protein and with a high affinity (K(D) = 4.7 ± 2 nM). The purified receptor appears to retain the pharmacological properties of the receptor in the original membranes. The purification resulted in over 80 % recovery of the initial ryanodine-binding sites and about 30-96-fold purification. This simple and fast method is highly reproducible and suitable for purification of small as well as large quantities of ryanodine receptor.

Original languageEnglish
Pages (from-to)61-64
Number of pages4
JournalBiochemical Journal
Volume285
Issue number1
DOIs
StatePublished - 1 Jan 1992

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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