Cation-exchange fast protein liquid chromatography – electrothermal atomic absorption spectrometry (FPLC – ETAAS) was applied for the speciation of trace amounts of aluminium. An aqueous 8 mol dm–3 NH4NO3 linear gradient elution in 10 minutes enabled separation of positively charged monomeric aluminium species in aqueous solution. Separated aluminium species were collected in 0.5 cm3 fractions ...

Neocarzinostatin, a highly toxic antitumor protein containing an essential nonprotein chromophore, can be isolated and purified from culture filtrates of Streptomyces carcinostaticus. Usually a lengthy procedure of up to 60 h is necessary for the isolation, including several chromatographic steps partly under conditions which favour inactivation of the drug by release of chromophore. We describ...

In this section, we provide the derivation of equation (3) (Main text). The objective of this calculation is to find an equation that defines CE(j). The equation should provide a value of # CE that is independent of i and that leads to a linear increase in the log10(D) in the progression from (imin; jmin) to (imin; jmax). We note that for all other values of i, the increase in log10(D) will the...

A dimeric 62-kDa lectin exhibiting a novel N-terminal amino acid sequence was purified from caper (Capparis spinosa) seeds. The purification protocol involved anion-exchange chromatography, cation-exchange chromatography and, finally, gel filtration by FPLC on Superdex 75. Approx. 100-fold purification was achieved. The haemagglutinating activity of the lectin, which was stable in the pH range ...

The cholera toxin (CT)-binding activity of purified κ-casein macropeptide (CMP) from bovine κ-casein was detected. In addition, a statistical model was developed to optimize the production of CMP. CMP was prepared by chymosin hydrolysis of κ-casein and a subsequent 3% trichloroacetic acid treatment. CMP was further fractionated in an ion-exchange column by FPLC. CT binding activity was eluted a...

Acetyl-CoA carboxylase (ACC), the first committed enzyme in fatty acid (FA) synthesis, is regulated by phosphorylation/dephosphorylation, transcription, and an unusual mechanism of protein polymerization. Polymerization of ACC increases enzymatic activity and is induced in vitro by supraphysiological concentrations of citrate (> 5 mM). Here, we show that MIG12, a 22 kDa cytosolic protein of pre...

Mitochondria of Plasmodium falciparum (K1 strain) were isolated by differential centrifugation. Mitochondrial DNA topoisomerase II from P. falciparum was partially purified using fast protein liquid chromatography(FPLC). Parasite mitochondria contained approximately 8% of DNA topoisomerase II activity compared with its nuclear fraction. The effects of fluoroquinolones, inhibitors of bacterial D...

Equinatoxin II (EqtII), a basic protein of 179 amino acids lacking cysteine residues, is the most abundant cytolysin isolated from the sea anemone Actinia equina. Its mode of action is still poorly understood. In order to initiate further structure-function studies by protein engineering, cDNA library was prepared from the whole animal and hybridized with a PCR-derived probe, deduced from the E...

Exactly why N-acetyl-beta-D-glucosaminidase (NAG) excretion is increased in patients with nephrotic syndrome with glomerular lesions is poorly understood. Glomeruli contain less NAG than do proximal tubules. In this study, we have tried to measure the NAG isoenzymes automatically by use of the recently developed fast protein liquid chromatography (FPLC) system, followed by column chromatography...