In vitro reconstitution and biophysical characterization of OEP16, an outer envelope pore protein of pea chloroplasts

D. Linke, J. Frank, J. F. Holzwarth, J. Soll, C. Boettcher, Petra Fromme

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

More than 30% of all proteins in the living cell are membrane proteins; most of them occur in the native membranes only in very low amounts, which hinders their functional and structural investigation. Here we describe the in vitro reconstitution of overexpressed Outer Envelope Protein 16 (OEP16) from pea chloroplasts, a cation-selective channel, which has been purified from E. coli inclusion bodies. Reconstitution in detergent micelles was monitored by CD and fluorescence spectroscopy. Electron microscopy showed a homogeneous size distribution of the reconstituted protein, and differential scanning calorimetry gave an estimate of the enthalpy of protein folding. First protein crystals were obtained that have to be further refined for X-ray structural analysis. The described methods of membrane protein reconstitution and biophysical analysis might prove helpful in the study of other membrane proteins.

Original languageEnglish
Pages (from-to)11050-11056
Number of pages7
JournalBiochemistry
Volume39
Issue number36
DOIs
Publication statusPublished - Sep 12 2000

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Porins
Peas
Chloroplasts
Membrane Proteins
Proteins
Protein folding
Fluorescence Spectrometry
Inclusion Bodies
Protein Folding
Differential Scanning Calorimetry
Fluorescence spectroscopy
Micelles
Structural analysis
Detergents
Escherichia coli
Electron microscopy
Cations
Differential scanning calorimetry
Enthalpy
Electron Microscopy

ASJC Scopus subject areas

  • Biochemistry

Cite this

In vitro reconstitution and biophysical characterization of OEP16, an outer envelope pore protein of pea chloroplasts. / Linke, D.; Frank, J.; Holzwarth, J. F.; Soll, J.; Boettcher, C.; Fromme, Petra.

In: Biochemistry, Vol. 39, No. 36, 12.09.2000, p. 11050-11056.

Research output: Contribution to journalArticle

Linke, D. ; Frank, J. ; Holzwarth, J. F. ; Soll, J. ; Boettcher, C. ; Fromme, Petra. / In vitro reconstitution and biophysical characterization of OEP16, an outer envelope pore protein of pea chloroplasts. In: Biochemistry. 2000 ; Vol. 39, No. 36. pp. 11050-11056.
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