Figure 3.

In vitro C-mannosylation reaction. (A) The peptide N-AC-KPPQFAWAQWFE-NH₂ (900 μM) was incubated with Dol-P-[2-³H]Man (0.25 μCi) and rat liver microsomes (145.8 μg of protein) for 30 min at 37°C or 2h at 26°C. After extraction with chloroform/methanol (3/2 vol/vol), the total amount of radioactivity in the aqueous phase, which contains the peptide, was determined. (B) Various polypeptides were incubated and analyzed as under ‘A’, except that the concentration of r-RNase 2 was 300 μM. The values were corrected for background by subtracting the value obtained in the absence of peptide. The data in A and B represent the average of two independent experiments.