Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Functional Complementation of an E. coli G-3-P Auxotrophic Mutant by GPDHc1. Expression construct pQE60+GPDHc1 and control vector pQE60 were introduced into E. colistrain BB20-14 harboring a lacZ suppressor plasmid pRE34. Single transformation colonies were inoculated onto M9 basal medium containing 10 mM glycerol, and media without glycerol but supplemented with various concentrations of IPTG.
• Supplemental Figure 2 - Sequence alignment of GPDHc1 with confirmed GPDHs from other organisms The amino acid sequences shown are (from top to bottom): GPDHc1, human cytosolic GPDH (H-GPDH, P21695), rat cytosolic GPDH (R-GPDH, NP_071551), E. coil gpsA (E-GPDH, AAB18585), and yeast cytosolic GPDH (Y-GPDH, NP_010262). Conserved residues are highlighted in yellow, and invariant residues are boxed in red. The residues that form the active sites based on crystal structural study and site directed mutagenesis are marked with blue asterisk (Choe et al., 2003). The NAD-binding domain GxGxxG (Wierenga et al., 1986) observed in all GPDHs studied to date is underlined.
• Supplemental Figure 3 - Restriction Mapping of the gpdhc1 Mutants. Left, blots of restricted genomic DNA of WS and gpdhc1-1 hybridized with theGPDHc1-specific probe and the T-DNA-specific BARprobe. Right,blots of restricted genomic DNA of Col and gpdhc1-2 hybridized with the GPDHc1-specific probe and the T-DNA-specific NPTII probe. The arrow shown next to each set of blots refers to the fragment that is detected by both the GPDHc1-specific probe and T-DNA-specific probe, due to T-DNA interruption in the mutant.
• Supplemental Table 1
• Supplemental Table 2
• Supplemental Methods