Title: The signal sequence of the p62 protein of Semliki Forest virus is involved in initiation but not in completing chain translocation Document date: 1990_9_1
ID: rmjv56ia_30
Snippet: Similar studies as with pCp62dhfr were also performed with the pCp62globin coded proteins in vitro. The results (not shown) were analogous to those described above for the pCp62dhfr construct. C protein and p62-globin hybrid were synthesized in the absence of membranes. When membranes were added, a protease-protected form of the hybrid appeared. This hybrid was also glycosylated as deduced from an experiment involving the acceptor peptide for gly.....
Document: Similar studies as with pCp62dhfr were also performed with the pCp62globin coded proteins in vitro. The results (not shown) were analogous to those described above for the pCp62dhfr construct. C protein and p62-globin hybrid were synthesized in the absence of membranes. When membranes were added, a protease-protected form of the hybrid appeared. This hybrid was also glycosylated as deduced from an experiment involving the acceptor peptide for glycosylation. Fig. 3 (lanes 1-6) shows the results of analyses in which we have tested whether the p62 signal sequence region confers stable membrane attachment to the p62-dhfr hybrid. Microsome-supplemented translations were adjusted to pH 11-11.5 with NaOH, incubated on ice for 10 min, and then separated into a membrane pellet and supernatant fraction by ultracentrifugation. In all samples the p62-dhfr polypeptides were isolated using an anti-dhfr antibody, SDS-PAGE shows that the hybrid protein segregates almost quantitatively into the supernatant fraction (compare lane I with lane 2). In similar conditions an integral membrane protein, the human transferrin receptor, was found to sediment with the membranes into the pellet fraction and a secretory protein, Ig light chain, was only recovered in the supernatant (not shown). If the acceptor peptide for glycosylation was included in the in vitro translation and the mixture then analyzed we found that the now unglycosylated but still translocated p62-hdfr hybrids were again mostly found in the supernatant fraction (lanes 3 and 4) . Lanes 5 and 6 show the analyses with the control peptide.
Search related documents:
Co phrase search for related documents- acceptor peptide and hybrid protein: 1, 2
- control peptide and hybrid protein: 1
Co phrase search for related documents, hyperlinks ordered by date