Anti-ACO1 | Aconitase

Aconitaseis 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

ArabidopsisACO1 (AT4G35830,Q42560), codon 120 – 898 (C-terminus), was cloned in fusion with a N-terminal 6xHis tag, and over-expressed inE. 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.

1 : 5 000 -1 : 10 000 (WB), At higher concentrations the antibody binds aspecifically resulting in non-specific signals around 60 kDa, including Rubisco subunits

The antibody recognises all three Arabidopsis aconitase isoforms (ACO1, ACO2 and ACO3, see Bernard et al 2009), Possible differences in affinity have not been precisely quantified, Sensitivity threshold is between 2 and 10 ng for WB / ECL (see figure), Antibodies will recognize aconitase isoforms in denaturing and native gel electrophoresis

Pascualet al (2021). ACONITASE 3 is part of the ANAC017 transcription factor-dependent mitochondrial dysfunction response, Plant Physiology, 2021;, kiab225, https://doi.org/10.1093/plphys/kiab225Przybyla-Toscanoet al. (2021) Protein lipoylation in mitochondria requires Fe-S cluster assembly factors NFU4 and NFU5. Plant Physiol. 2021 Oct 28:kiab501. doi: 10.1093/plphys/kiab501. Epub ahead of print. PMID: 34718778.Rureket al. (2018). Mitochondrial Biogenesis in Diverse Cauliflower Cultivars under Mild and Severe Drought Involves Impaired Coordination of Transcriptomic and Proteomic Response and Regulation of Various Multifunctional Proteins. Preprints 2018, 2018010276 (doi: 10.20944/preprints201801.0276.v1).Setiénet al. (2014). Root phosphoenolpyruvate carboxylase and NAD-malic enzymes activity increase the ammonium-assimilating capacity in tomato. J Plant Physiol. 171:49-63.Birkeet al. (2012). Cysteine biosynthesis, in concert with a novel mechanism, contributes to sulfide detoxification in mitochondria of Arabidopsis thaliana. Biochem J. May 2, ahead of print.

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.