Transport-specific fractionation for purification of ATP-dependent Ca2+ pumps

Stanley M. Goldin, Steven C. King

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

This chapter discusses transport-specific fractionation, which is a means for the identification of membrane transport proteins and as a purification procedure. The method depends on a specific biological property of a protein for isolation and employs a transport activity of the protein (rather than a ligand-binding property as would be used in affinity chromatography) as a physical tool for its purification. The most substantial success of this method in terms of a degree of purification has been its application by several laboratories to the purification of Ca2+-transport proteins, notably ATP-dependent Ca2+ pumps and the Na+/Ca2+ exchanger. The chapter discusses the methodology for the purification of reconstituted ATP-dependent Ca2+ pumps from mammalian brain synaptosomes. Two critical variables to consider when solubilizing the Ca2+-ATPase for use in transport-specific fractionation are the phospholipid-to-protein ratio and the cholate-to-phospholipid ratio.

Original languageEnglish (US)
Pages (from-to)34-49
Number of pages16
JournalMethods in Enzymology
Volume172
Issue numberC
DOIs
StatePublished - Jan 1 1989

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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