Journal Article

Yeast cell-based analysis of human lactate dehydrogenase isoforms

Lulu Ahmed Mohamed, Hiroyuki Tachikawa, Xiao-Dong Gao and Hideki Nakanishi

in The Journal of Biochemistry

Volume 158, issue 6, pages 467-476
Published in print December 2015 | ISSN: 0021-924X
Published online June 2015 | e-ISSN: 1756-2651 | DOI:
Yeast cell-based analysis of human lactate dehydrogenase isoforms

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  • Cell Biology
  • Biotechnology
  • Biochemistry
  • Molecular and Cell Biology


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Human lactate dehydrogenase (LDH) has attracted attention as a potential target for cancer therapy and contraception. In this study, we reconstituted human lactic acid fermentation in Saccharomyces cerevisiae, with the goal of constructing a yeast cell-based LDH assay system. pdc null mutant yeast (mutated in the endogenous pyruvate decarboxylase genes) are unable to perform alcoholic fermentation; when grown in the presence of an electron transport chain inhibitor, pdc null strains exhibit a growth defect. We found that introduction of the human gene encoding LDHA complemented the pdc growth defect; this complementation depended on LDHA catalytic activity. Similarly, introduction of the human LDHC complemented the pdc growth defect, even though LDHC did not generate lactate at the levels seen with LDHA. In contrast, the human LDHB did not complement the yeast pdc null mutant, although LDHB did generate lactate in yeast cells. Expression of LDHB as a red fluorescent protein (RFP) fusion yielded blebs in yeast, whereas LDHA-RFP and LDHC-RFP fusion proteins exhibited cytosolic distribution. Thus, LDHB exhibits several unique features when expressed in yeast cells. Because yeast cells are amenable to genetic analysis and cell-based high-throughput screening, our pdc/LDH strains are expected to be of use for versatile analyses of human LDH.

Keywords: alcoholic fermentation; lactate dehydrogenase; lactic acid fermentation; pyruvate decarboxylase; S. cerevisiae

Journal Article.  5733 words.  Illustrated.

Subjects: Cell Biology ; Biotechnology ; Biochemistry ; Molecular and Cell Biology

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