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. 2013 Nov 1;6(6):e27216.
doi: 10.4161/cib.27216. Epub 2013 Dec 10.

Fructose-1,6-bisphosphatase, Malate Dehydrogenase, Isocitrate Lyase, Phosphoenolpyruvate Carboxykinase, Glyceraldehyde-3-phosphate Dehydrogenase, and Cyclophilin A are secreted in Saccharomyces cerevisiae grown in low glucose

Bennett J Giardina  1 , Hui-Ling Chiang  1
Affiliations

Fructose-1,6-bisphosphatase, Malate Dehydrogenase, Isocitrate Lyase, Phosphoenolpyruvate Carboxykinase, Glyceraldehyde-3-phosphate Dehydrogenase, and Cyclophilin A are secreted in Saccharomyces cerevisiae grown in low glucose

Bennett J Giardina et al. Commun Integr Biol. .

Abstract

Our previous studies demonstrated that the key gluconeogenic enzyme fructose-1,6-bisphosphatase is secreted when Saccharomyces cerevisiae are starved of glucose for a prolonged period of time. In this study, we showed that malate dehydrogenase, isocitrate lyase, phosphoenolpyruvate carboxykinase, glyceraldehyde-3-phosphate dehydrogenase, and cyclophilin A are also secreted in glucose-starved cells. Thus, both gluconeogenic and non-gluconeogenic enzymes are secreted via the non-classical pathway.

Keywords: extracellular proteins; fructose-1,6-bisphosphatase; gluconeogenesis; malate dehydrogenase; non-classical secretion; periplasm; phosphoenolpyruvate carboxykinase; vacuole; vacuole import and degradation.

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Figures

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Figure 1. Immuno-TEM analysis of the distribution of FBPase, MDH2, Icl1p, Pck1p, GAPDH, and Cpr1p in glucose-starved wild-type cells. (A), wild-type cells expressing Icl1p-HA and Pck1p-Myc were lysed and total lysates were examined by western blotting using antibodies directed against Sec28p, FBPase, MDH2, HA, Myc, GAPDH, and Cpr1p. (B), wild-type cells expressing Icl1p-HA and Pck1p-Myc were starved of glucose for 3 d and fixed. Cells were processed and embedded. Thin sections (10 nm) were incubated in the absence or presence of primary antibodies directed against Sec28p, FBPase, MDH2, HA, Myc, GAPDH, and Cpr1p followed by secondary antibodies conjugated with 10 nm gold particles. Enlargements of the periplasm in sections of the whole cells were shown. CW:cell wall. PM:plasma membrane. Bars: 200 nm.
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Figure 2. . FBPase, MDH2, Icl1p, Pck1p, GAPDH, and Cpr1p are in the extracellular fraction in glucose-starved cells. Wild-type cells expressing Scw4p-GFP and wild-type cells co-expressing Icl1p-HA and Pck1p-Myc were grown in YPKG for three days and subjected to the extraction protocol. The distribution of Scw4p-GFP in the intracellular and extracellular fractions was examined by anti-GFP antibodies. The distribution of Sec28p, FBPase, MDH2, Icl1p-HA, Pck1p-Myc, GAPDH, and Cpr1p in the intracellular and extracellular fractions was examined using western blotting.
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