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Generation, characterization and crystallization of a highly active and stable cytochrome bc1 complex mutant from Rhodobacter sphaeroides.

Publication ,  Journal Article
Elberry, M; Xiao, K; Esser, L; Xia, D; Yu, L; Yu, C-A
Published in: Biochim Biophys Acta
July 2006

The availability of the three dimensional structure of mitochondrial enzyme, obtained by X-ray crystallography, allowed a significant progress in the understanding of the structure-function relation of the cytochrome bc(1) complex. Most of the structural information obtained has been confirmed by molecular genetic studies of the bacterial complex. Despite its small size and simple subunit composition, high quality crystals of the bacterial complex have been difficult to obtain and so far, only low resolution structural data has been reported. The low quality crystal observed is likely associated in part with the low activity and stability of the purified complex. To mitigate this problem, we recently engineered a mutant [S287R(cytb)/V135S(ISP)] from Rhodobacter sphaeroides to produce a highly active and more stable cytochrome bc(1) complex. The purified mutant complex shows a 40% increase in electron transfer activity as compared to that of the wild type enzyme. Differential scanning calorimetric study shows that the mutant is more stable than the wild type complex as indicated by a 4.3 degrees C increase in the thermo-denaturation temperature. Crystals formed from this mutant complex, in the presence of stigmatellin, diffract X-rays up to 2.9 Angstroms resolution.

Duke Scholars

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

July 2006

Volume

1757

Issue

7

Start / End Page

835 / 840

Location

Netherlands

Related Subject Headings

  • Rhodobacter sphaeroides
  • Protein Engineering
  • Electrophoresis, Polyacrylamide Gel
  • Electron Transport Complex III
  • Crystallography, X-Ray
  • Crystallization
  • Calorimetry, Differential Scanning
  • 51 Physical sciences
  • 31 Biological sciences
  • 06 Biological Sciences
 

Citation

APA
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Elberry, M., Xiao, K., Esser, L., Xia, D., Yu, L., & Yu, C.-A. (2006). Generation, characterization and crystallization of a highly active and stable cytochrome bc1 complex mutant from Rhodobacter sphaeroides. Biochim Biophys Acta, 1757(7), 835–840. https://doi.org/10.1016/j.bbabio.2006.05.031
Elberry, Maria, Kunhung Xiao, Lothar Esser, Di Xia, Linda Yu, and Chang-An Yu. “Generation, characterization and crystallization of a highly active and stable cytochrome bc1 complex mutant from Rhodobacter sphaeroides.Biochim Biophys Acta 1757, no. 7 (July 2006): 835–40. https://doi.org/10.1016/j.bbabio.2006.05.031.
Elberry M, Xiao K, Esser L, Xia D, Yu L, Yu C-A. Generation, characterization and crystallization of a highly active and stable cytochrome bc1 complex mutant from Rhodobacter sphaeroides. Biochim Biophys Acta. 2006 Jul;1757(7):835–40.
Elberry, Maria, et al. “Generation, characterization and crystallization of a highly active and stable cytochrome bc1 complex mutant from Rhodobacter sphaeroides.Biochim Biophys Acta, vol. 1757, no. 7, July 2006, pp. 835–40. Pubmed, doi:10.1016/j.bbabio.2006.05.031.
Elberry M, Xiao K, Esser L, Xia D, Yu L, Yu C-A. Generation, characterization and crystallization of a highly active and stable cytochrome bc1 complex mutant from Rhodobacter sphaeroides. Biochim Biophys Acta. 2006 Jul;1757(7):835–840.

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

July 2006

Volume

1757

Issue

7

Start / End Page

835 / 840

Location

Netherlands

Related Subject Headings

  • Rhodobacter sphaeroides
  • Protein Engineering
  • Electrophoresis, Polyacrylamide Gel
  • Electron Transport Complex III
  • Crystallography, X-Ray
  • Crystallization
  • Calorimetry, Differential Scanning
  • 51 Physical sciences
  • 31 Biological sciences
  • 06 Biological Sciences