**Appendix A**


**Figure A1.** Multiple sequence alignment of *Arabidopsis thaliana* pyruvate kinases.


**Figure A2.** Percentages of identity between *Arabidopsis thaliana* PK isoenzymes based on multiple sequence alignments of the respective amino acid sequences. The percentage of identity is shown in numbers and visualized by a green color gradient.

**Figure A3.** Histochemical GUS localization in oldest rosette leaves for the *cPK1* promoter (**A**,**D**,**G**), the *cPK2* promoter (**B**,**E**) and the *cPK3* promoter (**C**,**F**,**H**) in 3-week-old (**A**–**C**) and 5-week- old plants (**D**–**H**). Bars indicate 0.5 mm. G and H are magnifications of D and F, respectively. Representative images of one out of three different transgenic lines are shown.

**Figure A4.** Affinity purification of heterologous expressed cytosolic pyruvate kinases. Protein extracts (2 μL) of different fractions (1, cell extract; 2, flow through; 3, wash fraction I; 4, wash fraction II; 5-8, elution fraction I-IV) were separated on SDS-polyacrylamide gels and stained either by Coomassie protein stain or blotted and detected by Western blot analysis using anti-His antibodies.


**Table A1.** Oligonucleotide sequences for cloning.

**Figure A5.** Biochemical characterization of cytosolic pyruvate kinase isoenzymes. Km values of cPK1, cPK2, cPK3, cPK4 and cPK5 for ADP (left panel) and PEP (right panel) were determined by nonlinear regression based on the Michaelis–Menten equation. Linear dilutions of ADP (0.005, 0.01, 0.025, 0.05, 0.075, 0.1, 0.15, 0.25, 0.5, 2.5, 5.0 and 10.0 mM) and PEP (0.025, 0.05, 0.1, 0.25, 0.5 0, 0.75, 1.0, 1.25, 1.75, 2.5, 5.0 and 10.0 mM) were used. Data are mean ± SD of four replicates.
