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. 1972 Jan;109(1):385–390. doi: 10.1128/jb.109.1.385-390.1972

Purification and Characterization of Phosphoenolpyruvate Carboxylase from Plasmodium berghei

Huey G McDaniel a,1, Patrick M L Siu b
PMCID: PMC247289  PMID: 4621631

Abstract

Phosphoenolpyruvate (PEP) carboxylase was purified over 400-fold from Plasmodium berghei. The purified enzyme was stable in 0.4 m potassium phosphate buffer (pH 7.4) containing 0.5 m glucose, 1 mm ethylenediaminetetraacetic acid (EDTA), and 1 mm MgCl2. It had a molecular weight of 280,000 determined by sucrose density gradient centrifugation in this buffer, but it aggregated and was unstable in the presence of different salts or a more dilute solution of potassium phosphate. The Km for PEP was 2.6 mm and that for Mg2+ was 1.3 mm. The Km for bicarbonate was 2 mm. Citrate, nucleotides, and EDTA inhibited the PEP carboxylase of P. berghei by decreasing the concentration of free magnesium ions, but acetyl-coenzyme A, fructose-1,6-diphosphate, and aspartate did not influence its activity. A chloroquine concentration of 1.8 × 10−4m inhibited the enzyme 50%.

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Selected References

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