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Figure 5


Fig. 5. TM3 moves out of the ER translocon upon chain extension. (A) Membrane-associated integration intermediates of OP150 to OP259 containing a single cysteine probe in TM3 (residue 115) were treated with BMH and the resulting adducts analysed following immunoprecipitation as previously described. An adduct with a putative ribosomal protein of ~21 kDa was observed with OP150[Cys115] (panel A, lanes 2 and 3, labelled R) and in some cases adducts containing both Sec61{alpha} and Sec61ß were seen (labelled {alpha}ß). All other symbols as previously defined. (B) Membrane-associated integration intermediates of OP204 containing a single cysteine probe in TM4 (residue 154) were analysed as for panel A. A novel adduct was observed with the OP204[Cys154] intermediate (panel B, lanes 2 and 3, filled circle), this remains to be fully characterised. Trace levels of radiolabelled Sec61{alpha} resulting from the translation of endogenous mRNA that sometimes remained after the nuclease treatment of the semi-intact cells (data not shown) can be seen in panel B, lane 5 (X).





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