FIG 6.

Direct interaction of NCR247 with succinoglycan. Octet biolayer interferometry assays were performed as described in Materials and Methods. (A) Overview of all stages of an Octet biolayer interferometry assay. The steps include NCR247-biotin binding to the biosensor tip and the association of solubilized succinoglycan with immobilized NCR247-biotin. In a final step, the loaded biosensor tips are immersed in assay buffer without succinoglycan. (B to D) Octet biolayer interferometry assay results. The concentration of free succinoglycan was kept constant at 10 μg ml⁻¹ for the HMW and LMW forms. Where indicated, the ammonium acetate assay buffer was supplemented with 0.1 M NaCl. Times of 1.7 to 2.7 min, buffer; 2.7 to 8.2 min, buffer plus HMW or LMW succinoglycan; 8.2 to 10 min, buffer without salt (NaCl −) (B and C) or with 0.1 M salt (NaCl +) (D) (times varied minimally between experiments). Results shown are representative of trends observed in at least two independent experiments. wt, wild-type; EPS, exopolysaccharide (succinoglycan).