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Proteins, Expression, Isolation and Analysis | ... 2013). The cells were harvested and resuspended in 100 ml lysis buffer (25 mM Tris pH 7.9, 100 mM NaCl, and 1 mM DTT) for breakage. The supernatant obtained after centrifugation was incubated for 1 h at 4 °C with 20 ml of glutathione affinity resin (Genscript, USA). ... | Get A Quote |
The green alga Chlamydomonas reinhardtii contains six plastidic [2Fe2S]-cluster ferredoxins (FDXs), with FDX1 as the predominant isoform under photoautotrophic growth. FDX2 is highly similar to FDX1 and has been shown to interact with specific enzymes (such as nitrite reductase), as well as to share interactors with FDX1, such as the hydrogenases (HYDA), ferredoxin:NAD(P) reductase I (FNR1), and pyruvate:ferredoxin oxidoreductase (PFR1), albeit performing at low catalytic rates. Here we report the FDX2 crystal structure solved at 1.18 Å resolution. Based on differences between the Chlorella fusca FDX1 and C. reinhardtii FDX2 structures, we generated and purified point-mutated versions of the FDX2 protein and ... More