Pact suite and jcsg suite

Crystallization screenings were performed by high-throughput techniques in a NanoDrop robot and Innovadyne SD-2 microplates (Innovadyne Technologies Inc.), screening PACT Suite and JCSG Suite (Qiagen), JBScreen Classic 1–4 and 6 (Jena Bioscience) and Crystal Screen, Crystal Screen 2 and Index HT (Hampton Research). The conditions that produced crystals were optimized by sitting-drop vapor-diffusion method at 291 K by mixing 1 μL of protein solution and 1 μL of precipitant solution, equilibrated against 150 μL of precipitant solution in the reservoir chamber. The best crystals were obtained in a crystallization condition containing 0.15 M NaF and 16% (w/v) PEG3350 (Supplementary Fig. 1B). Protein concentration was assayed at the concentration range of 7–8 mg/mL.

Crystallization screenings were performed using high-throughput techniques in a NanoDrop robot and Innovadyne SD-2 microplates (Innovadyne Technologies, Inc.) with screening PACT Suite and JCSG Suite (Qiagen), JBScreen Classic 1 to 4 (Jena Bioscience), Morpheus and MIDASplus (Molecular Dimensions) and Crystal Screen I&II, SaltRX and Index HT (Hampton Research). The conditions that produced crystals were optimized with a sitting-drop vapor-diffusion method at 291 K by mixing 1μl of protein solution and 1 μl of precipitant solution, equilibrated against 150 μl of precipitant solution in the reservoir chamber. For p6CΔ31, the best crystals were obtained in a crystallization condition containing 0.1 M Bis–Tris Propane pH 7.0 and 1.3 M di-Ammonium Tartrate (Supplementary Figure S2B). For p6CΔ20, the best crystals were obtained in a crystallization condition containing 10% w/v PEG 8000; 20% v/v ethylene glycol; 0.02 M 1,6-hexanediol; 0.02 M 1-butanol; 0.02 M (RS)-1,2-propanediol; 0.02 M 2-propanol; 0.02 M 1,4-butanediol; 0.02 M 1,3-propanediol and 0.1 M MES/imidazole pH 6.5. In both cases, protein concentration was assayed at the concentration of 9–10 mg/ml.

Prior to setting up crystallization assays, CbpL buffer was exchanged to 10 mM Tris-HCl pH 7.5 and 140 mM choline chloride by dialysis. First crystallization screenings were performed by high-throughput techniques in a NanoDrop robot and Innovadyne SD-2 microplates (Innovadyne Technologies Inc.), screening PACT Suite and JCSG Suite (Qiagen), JBScreen Classic 1–4 and 6 (Jena Bioscience) and Crystal Screen, Crystal Screen 2 and Index HT (Hampton Research). Positive conditions were optimized by hanging-drop vapour diffusion method by mixing 1 μL of protein solution and 1 μL of precipitant solution, equilibrated against 500 μL of precipitant solution. Best crystals were obtained in a crystallization condition containing 1.6 M ammonium sulphate and 0.5 M LiCl, at a protein concentration of 40.8 mg/mL.