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Ssa1p cannot stably interact with the lumenal domain of Sec63p. GST–63Jp was prebound to glutathione–cross-linked agarose before Ssa1p and ATP were added, as described in MATERIALS AND METHODS. Proteins remaining in the supernatant (S) and bound to the pellet (P) were resolved by SDS-PAGE and visualized by Coomassie brilliant blue staining. Molecular weight markers are shown (×10−3) to the left (in kilodaltons).
