Figure 3.

Solubility and Traffic of Zeolin.

Leaf protoplasts from transgenic tobacco expressing zeolin were pulse-labeled with [³⁵S]Met and [³⁵S]Cys for 1 h and subjected to chase for the indicated times. Analysis was by SDS-PAGE and fluorography. The positions of zeolin (arrowheads), the 45-kD form (arrow), phaseolin fragments (vertical bars), the 95-kD putative O-glycosylated zeolin (asterisk), and contaminant polypeptides (dots) are indicated. Numbers at left indicate the positions of molecular mass markers in kD.

(A) Total homogenates were prepared from protoplasts, using homogenation buffer without 2-ME, and immunoselected using anti-phaseolin antiserum.

(B) Total homogenates were prepared from protoplasts or incubation medium and immunoselected with anti-phaseolin antiserum, using homogenation and immunoprecipitation buffers supplemented with 2-ME.

(C) Longer exposure of the fluorograph shown in (B).