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. 2022 Sep 29;18(9):e1010792. doi: 10.1371/journal.ppat.1010792

Fig 2. Binding of OsRMC to CBM1 causes to inhibit catalytic activity.

Fig 2

(A) MoCel10A-His was incubated with cellulose in the presence of a protein mixture (total 2.0 μg) of OsRMC-His (0–2.0 μg) and BSA (2.0–0 μg) for 1 h at 4°C, and proteins bound and unbound to cellulose were detected by immunoblot analysis using anti-His antibody. (B) MoCel10A-His (2.0 μg) was preincubated with a protein mixture (total 2.0 μg) of OsRMC-His (0–2.0 μg) and BSA (2.0–0 μg) for 30 min 4°C, and the hydrolytic activity towards a wheat cell wall preparation was determined. Data are means ± SD of three independent determinations. (C) Heat-inactivated wheat coleoptile segments were treated with sodium phosphate buffer (100 mM, pH 6.0) containing MoCel10A-His (2.0 μg) with BSA (2.0 μg) or a mixture of MoCel10A-His and OsRMC-His (2.0 μg each). Samples treated with buffer were used as a control. Extension measurement was performed by loading a constant 200 mN to approximately 5-mm segments for 3 min in an extensometer. Strain was calculated as (Lt−L0.1)/L0.1 (Lt, length of coleoptiles at each time point; L0.1, length of segments at time 0.1). Data are means ± SD of five independent determinations.