Purification and two-dimensional crystallization of highly active cytochrome b(6)f complex from spinach.
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.
Dietrich
J
J
Max-Planck-Institute of Biophysics, Department of Structural Biology, Heinrich-Hoffmann-Strasse 7, 60528, Frankfurt am Main, Germany.
Kühlbrandt
W
W
D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000737:chemistry; D003461:Crystallography; D003573:Cytochrome b Group / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D045346:Cytochrome b6f Complex; D004579:Electron Transport; D008854:Microscopy, Electron; D013053:Spectrophotometry; D018724:Spinacia oleracea / Q000737:chemistry*; D013997:Time Factors
D003573:Cytochrome b Group; D045346:Cytochrome b6f Complex