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p97 effects are influenced by TM segment composition. (A) Topology and methionine distribution of TM1–2 constructs. Location of residues Glu92 and Lys95 is shown within the TM1 sequence. (B) Carbonate extraction of wild-type and E92A/K95A polypeptides. (C, E) Degradation assays performed in mock-depleted RRL and p97-depleted RRL with or without added recombinant p97. Effects of MG132 and ATP depletion are also shown. (D, F) Initial degradation rates calculated from panels C and E.
