α d glucose 1 phosphate

TEV-cleavage experiments were performed using a PrfA* strain of L. monocytogenes that constitutively expresses ActA^{47 (link)}. These strains were grown for 12–16 h in 1.5X LB medium containing 7.5 μg mL⁻¹ chloramphenicol (Cm; Sigma) until reaching an OD₆₀₀=0.4–0.5. All other experiments were performed in the 10403S background. actA expression (10403S strains) from the native promoter^{7 (link)} or an IPTG-inducible promoter^{48 (link)} was induced as previously described. Bacteria were grown overnight in BHI medium containing 7.5 μg mL⁻¹ Cm, and then overnight cultures were spun down and resuspended in the appropriate medium. To express ActA from the SPAC/lacOid promoter, cultures were diluted 1:50 into BHI medium containing 7.5 μg mL⁻¹ Cm and 0.5 mM IPTG. To express ActA from its native promoter, cultures were diluted 1:25 into an induction medium of LB supplemented with 25 mM α-D-glucose 1-phosphate (Sigma) and ~1–2% (w/v) activated carbon pellets^{49 (link),50 (link)}.

The potassium salts of ADP and PEP were purchased from Chem-Impex International, Inc. NADH was from Sigma. L-lactate dehydrogenase (Type III bovine heart) was purchased from Calzyme Laboratories, Inc. Other buffer components were from Fisher Scientific and Sigma. The sodium salt of 2-deoxyribose 5-phosphate, the dipotassium salt of D-glucose 6-phosphate, The sodium salt of D-ribulose-1,5-bisphosphate, the dipotassium salt of α-D-glucose 1-phosphate and the disodium salt of D-fructose 6-phosphate were purchased from Sigma-Aldrich. DL-glyceraldehyde 3-phosphate, the sodium salt of D-fructose 1-phosphate, the dibarium salt of 2,5-anhydro-D-mannitol-1,6-bisphosphate, 2,5-anhydro-D-glucitol-1,6-bisphosphate and the cyclohexylammonium salt of α-D-glucose-1,6-bisphosphate were purchased from Santa Cruz Biotechnology. A table showing the structure of each analogue is included in the Supplemental Material.