Phase separation of rat intestinal brush border membrane proteins using Triton X-114

Chinnaswamy Tiruppathi, David H. Alpers, Bellur Seetharam

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Rat intestinal microvillus membrane contains at least 24 polypeptides, of which 18 can be solubilized using Triton X-114 at 4°C. Upon phase separation at 32°C, 11 proteins separated nearly completely into the detergent-rich phase, while 9 proteins were found exclusively in the aqueous phase. Enzymes which were uniquely included in the detergent phase were alkaline phosphatase, leucine aminopeptidase, γ-glutamyl transpeptidase, and Ca2+Mg2+ ATPase. The proteins which were excluded from the detergent phase and found exclusively in the aqueous phase included the disaccharidases (glucoamylase, sucrase-isomaltase, trehalase, lactase) and the ileal receptor for the intrinsic factor-cobalamin complex. Integral membrane proteins can thus be separated during solubilization into two groups prior to further purification or characterization.

Original languageEnglish
Pages (from-to)330-335
Number of pages6
JournalAnalytical Biochemistry
Volume153
Issue number2
DOIs
StatePublished - Mar 1986

Keywords

  • Triton X-114
  • electrophoresis
  • intestine
  • membrane
  • phase separation
  • proteins

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