Aconitase is a single subunit enzyme of the tricarboxylic acid cycle (or Krebs cycle) in the mitochondria. A cytosolic isoform is also part of the glyoxylate cycle. Aconitase catalyzes the dehydration / hydration of citrate to iso-citrate, via cis-aconitate as an intermediate. The reaction is facilitated by an iron-sulphur cluster in the active site of the enzyme. The iron-sulphur cluster is somewhat unstable, especially under oxidative stress, and loss of the cofactor leads to degradation of the protein.Alternative names: ACO, citrate hydro-lyase 1,2,3
應(yīng)用類型
Western blot (WB)
免疫原
Arabidopsis ACO1 (AT4G35830, Q42560), codon 120 - 898 (C-terminus), was cloned in fusion with a N-terminal 6xHis tag, and over-expressed in E. coli. All recombinant protein accumulated in inclusion bodies, which were purified by centrifugation and solubilised in 6 M guanidine-HCl. The protein was refolded by dilution in 100 mM Tris-HCl 8.5, 10% (v/v) glycerol, 2 mM dithiothreitol, and concentrated prior to immunisation.
來源宿主
兔
反應(yīng)性
擬南芥 ACO1,ACO2 and ACO3 isoforms, Brassica oleracea, Solanum lycopersicum