|Formulation:||In 50% (v/v) glycerol|
|Stability:||Minimum 1 year at < -10oC. Check vial for details.|
|Enzyme Activity:||Other Activities|
|EC Number:||184.108.40.206 (Modified from 220.127.116.11 in 2002)|
|Synonyms:||citrate (Si)-synthase; acetyl-CoA:oxaloacetate C-acetyltransferase [thioester-hydrolysing, (pro-S)-carboxymethyl forming]|
|Concentration:||Supplied at ~ 300 U/mL|
|Expression:||Recombinant from Escherichia coli|
Catalyses the reaction:
Acetyl-CoA + H2O + oxaloacetate = citrate + CoA
|Specific Activity:||~ 10 U/mg (25oC, pH 8.0 on oxaloacetic acid)|
|Unit Definition:||One Unit of citrate synthase is defined as the amount of enzyme required to produce one µmole of citric acid from oxaloacetic acid (0.16 mM) and acetyl-CoA (0.2 mM), measured at 232 nm in Tris.HCl buffer (95 mM), pH 8.0 at 25oC.|
|Application examples:||Applications for the measurement of acetic acid in the food, fermentation, wine, beverage and dairy industries.|
High purity recombinant Citrate synthase (Escherichia coli) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.
Discover more high purity enzyme products for analytical applications.
Mädler, S., Barylyuk, K., Erba, E. B., Nieckarz, R. J. & Zenobi, R. (2012). Journal of The American Society for Mass Spectrometry, 23(2), 213-224.
Chemical cross-linking in combination with high-mass MALDI mass spectrometry allows for the rapid identification of interactions and determination of the complex stoichiometry of noncovalent protein–protein interactions. As the molecular weight of these complexes increases, the fraction of multiply charged species typically increases. In the case of homomeric complexes, signals from multiply charged multimers overlap with singly charged subunits. Remarkably, spectra recorded in negative ion mode show lower abundances of multiply charged species, lower background, higher reproducibility, and, thus, overall cleaner spectra compared with positive ion mode spectra. In this work, a dedicated high-mass detector was applied for measuring high-mass proteins (up to 200 kDa) by negative ion mode MALDI-MS. The influences of sample preparation and instrumental parameters were carefully investigated. Relative signal integrals of multiply charged anions were relatively independent of any of the examined parameters and could thus be approximated easily for the spectra of cross-linked complexes. For example, the fraction of doubly charged anions signals overlapping with the signals of singly charged subunits could be more precisely estimated than in positive ion mode. Sinapinic acid was found to be an excellent matrix for the analysis of proteins and cross-linked protein complexes in both ion modes. Our results suggest that negative ion mode data of chemically cross-linked protein complexes are complementary to positive ion mode data and can in some cases represent the solution phase situation better than positive ion mode.Hide Abstract