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Purification and two‐dimensional crystallization of highly active cytochrome b 6 f complex from spinach
Author(s) -
Dietrich Jens,
Kühlbrandt Werner
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01609-9
Subject(s) - spinach , crystallization , dimethyl sulfoxide , yield (engineering) , solvent , chemistry , cytochrome , electron transfer , crystallography , chromatography , analytical chemistry (journal) , materials science , organic chemistry , biochemistry , enzyme , metallurgy
The purification and two‐dimensional crystallization of highly active cytochrome b 6 f complex from spinach is described. The preparation shows all spectroscopic characteristics of the pure complex. The electron transfer activity of 450±60 electrons per s is the highest in vitro activity reported to date. Using dimethyl sulfoxide (DMSO) as a solvent for the electron donor enhanced the performance and reproducibility of the assay. The high yield and the high activity of the protein make it an ideal candidate for biophysical and structural studies. Preliminary two‐dimensional crystallization experiments yielded several different forms of two‐dimensional and thin three‐dimensional crystals, exhibiting varying degrees of order.