Figure 6.

Substitution of Ero1p loop residues decreases recovery of an interdomain Cys105-Cys352 disulfide bond. Transformants of CKY598 (ero1-1) containing pCS253, pCS347, pCS349, pCS348, pCS306, pCS307, or pCS262 were radiolabeled at 38°C for 30 min, lysed in conditions to block disulfide exchange, and Ero1p was immunoprecipitated with anti-myc antibody under denaturing, nonreducing conditions. Immunoprecipitates were cleaved with thrombin and samples were resolved by SDS-PAGE under nonreducing conditions.