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. 2011 Apr 25;108(19):7775–7780. doi: 10.1073/pnas.1101705108

Fig. 5.

Fig. 5.

IsoTAM2-proOmpA is translocated by SecY with a lateral gate that is constrained by a 8.6 Å cross-linker. (A) Top view (Left) and side view (Right) of the crystal structure of the M. jannaschii SecYEβ. SecE and SecG are indicated in green and orange, respectively. TM 2 and 7 that form the lateral gate are indicated in blue. The red balls indicate the crosslinking sites. (B) OmpT assay performed on IMVs containing the cysteineless and SecY(F286C/S87C)EG complex incubated with different chemical cross linkers. In the presence of sodium tetrathionate (lane 4), or bis-maleimidoethane (lane 5), the OmpT-treated SecY migrates as the uncleaved protein (lane 2). In the presence of TCEP, SecY is cleaved (lane 3). The molecular mass standard is indicated in lane 1. (C) Translocation of Fmal-proOmpA (Left) and IsoTAM2-proOmpA (Right) into SecY(F286C/S87C)EG IMVs under reducing conditions (lanes 2 and 6), or upon treatment with sodium tetrathionate (lanes 3 and 7), or bis-maleimidoethane (lanes 4 and 8). As a control, no ATP was added to the translocation reaction (lanes 1 and 5).