Extended Data Fig. 1: Enzyme activity of plastid-localized ENO and PGM from isolated chloroplasts of Arabidopsis.
From: Rubisco supplies pyruvate for the 2-C-methyl-d-erythritol-4-phosphate pathway
Crude protein extracts were obtained from intact chloroplasts isolated from 7-day-old seedlings and rosette-staged leaves (42 days) and incubated with 2 mM 3-PGA or 1 mM 2-PGA to assess the relative activity of PGM and ENO glycolytic enzymes. a, Assay showing lower activity of plastidic ENO in rosette leaves when compared to seedlings. b, Coupled enzyme activity of PGM and ENO shows low conversion of 3-PGA to PEP in rosette leaves (ENO P = 0.0002; PGM + ENO P = 0.0026). c, Representative chromatograms depicting PEP product formation after one hour incubation with 2-PGA. All chromatograms are shown on the same scale. d, Intact chloroplasts isolated from 7-day-old seedlings. Bar = 10 μm. The experiment was repeated once with identical results. Error bars represent standard deviation of measurements (n = 3). Asterisks indicate significant differences (** P < 0.001; *** P < 0.0001) as determined by Student’s two-sided t-test.
