Bovine rnase a

SEC experiments were performed at pH 8.0 (50 mM Tris) and 0.250 M NaCl in a Superose 12 10/300 GL column, which was connected to an AKTA FPLC (GE Healthcare, Barcelona, Spain); absorbance at 280 nm was monitored during the elution. Flow rate was 0.8 mL/min, and C-LrtA was loaded in a volume of 100 μL at several protein concentrations, in the range of 30–500 μM (in protomer concentration). The markers used to calibrate the column were ferritin, catalase, aldolase, albumin, bovine RNase A and blue dextran (GE Healthcare, Barcelona, Spain); the isolated protein markers were loaded in the column in the above described buffer, at the same flow rate, and three independent measurements were performed with each marker.

SEC experiments were carried out in a Superose 12 10/300 GL column at pH 8.0 (50 mM Tris) and 0.7 M NaCl, connected to an AKTA FPLC (GE Healthcare), and monitoring the absorbance at 280 nm. Flow rate was 0.7 mL/min, and the protein was loaded in a volume of 100 μL at a concentration of 97 μM (in protomer units). Baselines of the chromatograms were strongly curved and therefore the UNICORN software 5.0.2 (GE Healthcare, Barcelona, Spain) was used for automatic baseline correction. The markers used to calibrate the column were albumin, bovine RNase A and blue dextran (GE Healthcare); the isolated markers were loaded in the column in the above described buffer in three independent measurements.