Figure 2.

A transient disulfide bond forms between Ero1p Cys105 and Cys352. Cells with a chromosomal deletion of ERO1 containing pCS253 (A) or transformants of CKY598 (ero1-1) containing pCS253, pCS257, pCS258, pCS259, pCS254, pCS255, or pCS256 (B) were radiolabeled for 30 min at 30°C (A) or 38°C (B), lysed in conditions to block disulfide exchange, and Ero1p was immunoprecipitated under denaturing, nonreducing conditions with monoclonal anti-myc. Immunoprecipitates were cleaved with thrombin and samples were resolved by SDS-PAGE under reducing or nonreducing conditions.