Figure 5.

Specific GNOM and GNOM DCB Domain Binding to Cyp5 in Vitro.

(A) GST–Cyp5 binding of GNOM protein from Arabidopsis extracts. Protein gel blot analysis detected a 165-kD full-length GNOM band with anti-GNOM Sec7 antiserum (Anti-GNOM). Controls are beads or GST-coupled beads (GST) incubated with plant extract.

(B) and (C) Binding of ³⁵S-methionine–labeled in vitro translation products to GST fusion proteins. Autoradiographs are of 15% SDS–polyacrylamide gels. In (B), specific binding of GNOM DCB domain (amino acids 1 to 246) to GST–Cyp5 is shown. Equal amounts of translation product were incubated with equal concentrations of GST, GST–ROC1, and GST–Cyp5. In (C), specific binding of Cyp5 translation product to GST–GNOM DCB domain (amino acids 1 to 246) is shown. Equal amounts of translation product (10% input) of ROC1 and Cyp5 (arrows) were incubated with equal amounts of GST and GST–GNOM amino acids 1 to 246.