PMID: 9445396Mar 7, 1998Paper

Cloning and expression of the gene for the active PPi-dependent phosphofructokinase of Entamoeba histolytica

The Biochemical Journal
Z DengR G Kemp

Abstract

Pyrophosphate-dependent phosphofructokinase (PPi-PFK) from Entamoeba histolytica (HM-1) was purified from trophozoites. Oligonucleotide probes based on partial amino acid sequence were used to clone and sequence the gene and the cDNA of the enzyme. The molecular mass of the subunit was greater than, and the derived sequence significantly different from, that of the product of the PPi-PFK gene previously cloned from E. histolytica [Huang, Albach, Chang, Tripathi and Kemp (1995) Biochim. Biophys. Acta 1260, 215-217; Bruchhaus, Jacobs, Denart and Tannich (1996) Biochem. J. 316, 57-63]. The sequence identity between the two proteins was 17%. The sequence bore greater identity with the more phylogenetically advanced plant PPi-PFKs than with bacterial PPi-PFKs. The cloned cDNA was expressed and the protein purified. The kinetic properties were identical with those of the enzyme isolated from the organism. Furthermore, the specific activity was more than three orders of magnitude higher than that described for the product of the previously cloned E. histolytica PFK gene [Bruchhaus et al. (1996)]. The pH-dependence and apparent substrate affinities of the cloned enzyme were identical with those of the PPi-PFK in trophozoite extracts, i...Continue Reading

Citations

Jan 31, 2008·Molecular Genetics and Genomics : MGG·Fabrício Ramon LopesClaudia Marcia Aparecida Carareto
Mar 23, 2001·The Journal of Biological Chemistry·A S ChiR G Kemp
May 31, 2008·The FEBS Journal·Rafael Moreno-SánchezEmma Saavedra
Sep 26, 2000·The Journal of Biological Chemistry·A Chi, R G Kemp
Sep 11, 2007·The FEBS Journal·Emma SaavedraRafael Moreno-Sánchez
Apr 25, 2013·Applied Biochemistry and Biotechnology·Rachel Ruizhen ChenZichao Mao
Oct 1, 1998·Biochemical and Biophysical Research Communications·X WangR G Kemp
Jun 10, 2009·International Journal for Parasitology·Binny M MonyShobhona Sharma

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