Figure 4.
Fractionation of ZM-INVINH1. A, Proteins isolated from 5-DAP (odd lanes) and 10-DAP (even lanes) kernels were fractionated into soluble (lanes 1 and 2), high-salt wash (lanes 3 and 4), and insoluble fractions (lanes 5 and 6) and subjected to western-blot analysis with antibody raised against ZM-INVINH1. Fifty micrograms of total soluble protein and 20 μg of high-salt and insoluble protein were separated by SDS-PAGE, and western-blot analysis was performed with ZM-INVINH1 antibody. B, ZM-INVINH1 associates with the glycoprotein fraction in maize kernels. Glycoproteins were isolated from 5- and 10-DAP kernels by ConA chromatography, and ZM-INVINH1 was detected by western-blot analysis. Lanes 1 and 2 correspond to ConA chromatography flow through and eluate, respectively. A large arrow represents ZM-INVINH1, and an asterisk indicates a high Mr cross-reacting protein.