Phenyl-sepharose-mediated detergent-exchange chromatography: Its application to exchange of detergents bound to membrane proteins

Neal C. Robinson, Diane Wiginton, Linda Talbert

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Detergent-saturated phenyl-Sepharose was used to exchange detergents for one another in the presence of membrane proteins. The alkyl detergents lauryl maltoside, octyl glucoside, and dodecyl sulfate were each successfully exchanged for Triton X-100, Triton N-101, or Nonidet P-40 present in a solution of either cytochrome c oxidase, a mixture of inner mitochondrial membrane proteins, or a mixture of erythrocyte membrane proteins. The method involves (1) saturating a small column of phenyl-Sepharose (1-2 mL) with one of the alkyl detergents at a pH of 8 or 9 and an ionic strength of 0.01, (2) applying a detergent-solubilized membrane protein sample containing as much as 20 mg/mL of Triton X-100, Triton N-101, or Nonidet P-40, and (3) eluting the protein with buffer containing the detergent with which the resin had been saturated. With this approach, 90-99% of the detergent in the initial protein sample was exchanged for the second detergent with an 80-100% recovery of protein. The advantages of this method over previous approaches for exchanging detergents include the rapidity of the technique and the apparent general applicability of the method to a wide variety of detergents and membrane proteins.

Original languageEnglish (US)
Pages (from-to)6121-6126
Number of pages6
JournalBiochemistry
Volume23
Issue number25
StatePublished - 1984

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Chromatography
Detergents
Membrane Proteins
Octoxynol
phenyl-sepharose
Proteins
Mitochondrial Proteins
Erythrocyte Membrane
Mitochondrial Membranes
Electron Transport Complex IV
Ionic strength
Osmolar Concentration
Buffers
Resins
Recovery

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phenyl-sepharose-mediated detergent-exchange chromatography : Its application to exchange of detergents bound to membrane proteins. / Robinson, Neal C.; Wiginton, Diane; Talbert, Linda.

In: Biochemistry, Vol. 23, No. 25, 1984, p. 6121-6126.

Research output: Contribution to journalArticle

Robinson, Neal C. ; Wiginton, Diane ; Talbert, Linda. / Phenyl-sepharose-mediated detergent-exchange chromatography : Its application to exchange of detergents bound to membrane proteins. In: Biochemistry. 1984 ; Vol. 23, No. 25. pp. 6121-6126.
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