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. 2003 Aug;132(4):2073–2085. doi: 10.1104/pp.103.020024

Figure 7.

Figure 7.

Residual wall-loosening activity after removal of unbound Zea m 1 and after protease treatment. A, After 30 min of extension of the heat-inactivated wall in 50 mm sodium acetate (pH 4.5), the incubation solution was changed for one containing 0.3 mg mL-1 Zea m 1d, and the wall was extended for a further 30 min. The solution in the extensometer cuvette was exchanged 5× with buffer without Zea m 1d (“Treatment”). The negative control contained no Zea m 1d, whereas the positive control continued extending in 0.3 mg mL-1 Zea m 1d. B, Destruction of wall extension activity by Pronase. Heat-inactivated wheat coleoptiles were pre-incubated for 1 h with 0.1 mg mL-1 of Zea m 1d in 50 mm sodium acetate, pH 4.5. After three washes with 50 mm MES, 1 mm EDTA, and 5 mm DTT (pH 6.0), they were then further pretreated for 1 h at room temperature with 2 mg mL-1 of Pronase in the washing buffer. The walls were then assayed for residual Zea m 1 activity. At the time indicated by the arrow, the buffer was switched from pH 7.5 buffer to pH 6.0 buffer (50 mm DMGA in both cases). All of the above experiments were performed at least five times with similar results.