Bioinformatic and Structural Comparison T6PP Orthologs

Root mean square deviation (RMSD) values of the α-carbons were calculated using the PDB Calculate Structure Alignment tool (23). The size of the active-site cavity was measured with Voidoo (24), and tunnels to/from the active site were determined using Caver (25). Molecular graphics were performed using UCSF Chimera (26) unless otherwise noted. UCSF Chimera was also used to determine hydrogen bonds (Find Hbond (27)) or van der Waals interactions (Find Contacts (28)) made between enzyme residues and ligand. Two-dimensional depictions were drawn in ChemDraw^® Professional 16. Multiple sequence alignments were generated using Clustal Omega (29) and visualized with ESPript 3 (30). For conservational analysis, 80 sequences of T6PP from bacteria, fungi, plants, insects and nematodes were included (see supplement for list of UniProt IDs). Each sequence is manually curated by UniProt based on sequence analysis and experimental evidence from literature (31). Sequence identities were extracted from a multiple sequence alignment of Bm, Ce, Cn, Af, Ca, Ta, St, Mt, and Mm T6PP sequences.