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Effect of mutations D743R and D744R on TM7b ST activity and TM8 topology. Topology of polypeptides generated from plasmids S.gG.TM7b(DD/RR).P (A), S.gG.TM7b(DD/RR)-8.P (B), and TM7a/b(DD/RR).P (C) was determined as described in Figure 2. Downward and upward arrows indicate full-length chains and PK-protected fragments, respectively. Topology of resulting chains is diagrammed, and the percent of polypeptides in the unconventional Pgp topology is indicated.
