New approach of three-dimensional crystallization of the Ca(2+)-ATPase of sarcoplasmic reticulum.

T. Keresztes, S. Varga, A. Kövér

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Abstract

Selective extraction procedure was applied for obtaining different proteins from sarcoplasmic reticulum vesicles following the main steps of previous observations. High concentration of a nonionic detergent, such as polyoxyethylene-10-lauryl ether (C12E10) prevented the formation of Ca(2+)-ATPase crystals. It has been observed that only the 300 kDa protein could induce crystallization from among proteins being undissolved from of membrane. A modification of MacLennan's procedure--applied for ATPase precipitation from deoxycholate solubilized sarcoplasmic reticulum--has been described and an ammonium acetate precipitated Ca(2+)-ATPase was used in the experiments for increase of Ca(2+)-ATPase concentration in the crystallization process. The repeated supplementation of purified and C12E10 solubilized Ca(2+)-ATPase with ammonium acetate precipitated Ca(2+)-ATPase made possible a formation of larger and larger crystals with different periodicity.

Original languageEnglish
Pages (from-to)125-132
Number of pages8
JournalActa biochimica et biophysica Hungarica
Volume25
Issue number1-2
Publication statusPublished - 1990

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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