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, P. Fromme

Research output: Contribution to journalArticle

9 Scopus citations

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 (US)
Pages (from-to)11050-11056
Number of pages7
JournalBiochemistry
Volume39
Issue number36
DOIs
StatePublished - Sep 12 2000
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'In vitro reconstitution and biophysical characterization of OEP16, an outer envelope pore protein of pea chloroplasts'. Together they form a unique fingerprint.

  • Cite this