Crystallization and preliminary structure of beef heart mitochondrial cytochrome-bc1 complex

Chang An Yu, Jia Zhi Xia, Anatoly M. Kachurin, Linda Yu, Di Xia, Hoeon Kim, Johann Deisenhofer

Research output: Contribution to journalArticle

83 Scopus citations

Abstract

The method reported for isolation of ubiquinol-cytochrome-c reductase complex from submitochondrial particles was modified to yield a preparation for crystallization. The cytochrome be, complex was first crystallized in large thin plate form and diffracts X-rays to 7 A resolution in the presence of mother liquor. This crystalline complex was enzymatically active and contains ten protein subunits. It had 33 mol phospholipid and 0.6 mol ubiquinone per mol protein. With slightly modified crystallization conditions, different crystal forms were obtained. Crystals grown in the presence of 20% glycerol diffracted X-rays up to 2.9 Å resolution using a synchrotron source. Four heavy atom derivatives have been obtained. The 3-D structure of the cytochrome bc1 complex was solved to 3.4 Å resolution. Crystalline cytochrome bc1 complex is a dimer: most of the masses of core proteins I and II protrudes from the matrix side of the membrane, whereas the cytochrome b protein is located mainly within the membrane. There are 13 transmembrane helices in each monomer. Most of the mass of cytochrome c1 and iron-sulfur protein including their redox centers are located on the cytoplasmic side of the membrane. The distances between these redox centers have been determined, and several electron transfer inhibitor binding sites in the complex have been located.

Original languageEnglish (US)
Pages (from-to)47-53
Number of pages7
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1275
Issue number1-2
DOIs
StatePublished - Jul 18 1996

Keywords

  • Co-crystallization of cytochrome bc-cytochrome c complex
  • Electron transport
  • Inhibitor complex
  • Mitochondrial cytochrome bc complex
  • Protein crystallization

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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