Solubilization of sarcoplasmic reticulum with Triton X-100

Bentson McFarland, G. Inesi

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The nonionic detergent Triton X-100 has been found to solubilize approximately 80% of the protein, and nearly all the phospholipid from sarcoplasmic reticulum membranes. The soluble fraction contains Ca2+ dependent ATPase, but no basic ATPase activity. In comparative experiments, Triton X-100 causes much less enzyme denaturation than ionic detergents. Analytical ultracentrifugation and disc electrophoresis indicate that the Triton-solubilized material consists of particles having a a molecular weight of approximately 80,000 daltons. These particles have a strong tendency to aggregate into size isomers. The aggregation proceeds at a faster rate in the presence of MgCl2, and is temperature dependent with an optimum at 25°. When the solubilized fraction is centrifuged on a sucrose density gradient, [32P]-labeled protein and phospholipid are separated, and ATPase activity is lost. If ATP is present in the density gradient during centrifugation, both ATPase inactivation and separation of protein from phospholipid are partially prevented. Gel electrophoresis of SR solubilized with sodium dodecylsulfate (SDS) shows that approximately 60% of the membrane protein migrates in a band corresponding to a molecular weight of 90,000 daltons. This fraction is absent in residues of SR previously extracted with Triton. Minor components are also separated in the SDS gels.

Original languageEnglish (US)
Pages (from-to)456-464
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume145
Issue number2
DOIs
StatePublished - 1971
Externally publishedYes

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Octoxynol
Sarcoplasmic Reticulum
Adenosine Triphosphatases
Phospholipids
Detergents
Molecular Weight
Electrophoresis
Gels
Sodium
Disc Electrophoresis
Proteins
Magnesium Chloride
Density Gradient Centrifugation
Calcium-Transporting ATPases
Molecular weight
Ultracentrifugation
Denaturation
Sucrose
Centrifugation
Membrane Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Solubilization of sarcoplasmic reticulum with Triton X-100. / McFarland, Bentson; Inesi, G.

In: Archives of Biochemistry and Biophysics, Vol. 145, No. 2, 1971, p. 456-464.

Research output: Contribution to journalArticle

McFarland, Bentson ; Inesi, G. / Solubilization of sarcoplasmic reticulum with Triton X-100. In: Archives of Biochemistry and Biophysics. 1971 ; Vol. 145, No. 2. pp. 456-464.
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