Over-production of lactate dehydrogenase from Plasmodium falciparum opens a route to new antimalarials

D Turgut-Balik, D K Shoemark, K M Moreton, R B Sessions, J J Holbrook

Research output: Contribution to journalArticle (Academic Journal)peer-review

19 Citations (Scopus)

Abstract

Over-production of lactate dehydrogenase (PfLDH) from Plasmodium falciparum from E. coli TG2 cells transformed with a pKK223-3 plasmid containing the wild type gene isolated by Bzik DJ, Fox BA, and Gonyer K (1993) Mol. Biochem. Parasit. 59, 155-166, gave mostly an inactive protein after isolation. Sequencing the N-terminus of the over-produced protein showed that the major product commenced at an internal methionine. Truncation of the protein occurred due to the inappropriate priming from a Shine-Dalgarno (SD) sequence upstream of Met 35. Silent mutations of this SD sequence to remove the purine-rich region allowed over-production of the full length PfLDH up to 15 mg protein l(-1) broth. The purified protein exhibited biochemical properties of an authentic LDH enzyme. However, high activity with 3-acetylpyridine adenine dinucleotide as well as with the natural cofactor, NAD, was also observed. The high-resolution X-ray structure obtained from the recombinant enzyme has provided the opportunity for the development of inhibitors specific to PfLDH.

Translated title of the contributionOver-production of lactate dehydrogenase from Plasmodium falciparum opens a route to new antimalarials
Original languageEnglish
Pages (from-to)917-921
Number of pages5
JournalBiotechnology Letters
Volume23
Issue number11
Publication statusPublished - Jun 2001

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