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Figure 2 | BMC Cell Biology

Figure 2

From: Different conformations of nascent polypeptides during translocation across the ER membrane

Figure 2

Determination of dP-O values for constructs where a model polypeptide segment has been inserted into the middle of the C-terminal P2 domain of Lep. (A) Two constructs with a ...QQQV18KKKK... model segment in the C-terminal domain were translated in vitro in the absence (-) and presence (+) of rough dog pancreas microsomes (RM). The 3' codon in the truncated mRNA was placed d codons downstream of the Asn residue in the Asn-Ser-Thr glycosylation acceptor site. The Asn residue in the acceptor site was placed in lep codon 178 (upper panel) or 200 (lower panel), and the model segment was inserted between lep codons 214 and 220 in both constructs. For the indicated range of d values, the model segment is located in the ribosome (upper panel) or in the translocon (lower panel). Non-glycosylated and glycosylated molecules are indicated by black and white dots, respectively. (B) Glycosylation profiles for constructs with model segments composed of the indicated residues flanked by QQQ....KKKK. The Asn residue in the acceptor site was placed in lep codon 178 (upper panel) or 200 (lower panel), and the model segment was inserted between lep codons 214 and 220 in all constructs. Half-maximal glycosylation (30%) is indicated by the horizontal line.

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