Abstract：
:To study the insertion of multispanning membrane proteins into the endoplasmic reticulum, we constructed novel proteins on the cDNA level by repeating, up to four times, the internal signal-anchor domain of the asialoglycoprotein receptor H1. Upon in vitro translation in the presence of microsomes, these polypeptides are indeed inserted as polytopic membrane proteins. The first hydrophobic domain functions as a signal and the second as a stop-transfer sequence, while the third initiates a second translocation process, halted again by the fourth. We were able to demonstrate that insertion occurs sequentially, starting with the first apolar segment from the amino terminus. By replacing the original signal-anchor domains by a mutant sequence not recognized by signal recognition particle (SRP), it was shown that only the first hydrophobic domain needs to be a signal sequence and that the second translocation event does not require SRP.
journal_name
Celljournal_title
Cellauthors
Wessels HP,Spiess Mdoi
10.1016/0092-8674(88)90009-8subject
Has Abstractpub_date
1988-10-07 00:00:00pages
61-70issue
1eissn
0092-8674issn
1097-4172pii
0092-8674(88)90009-8journal_volume
55pub_type
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