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PMID: 1848865 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Requirements for the membrane insertion of signal-anchor type proteins.

The Journal of cell biology ·Vol. 113 ·No. 1 ·1991-04-00 ·Pages 25-34

High S, Flint N, Dobberstein B

Abstract

Proteins which are inserted and anchored in the membrane of the ER by an uncleaved signal-anchor sequence can assume two final orientations. Type I signal-anchor proteins translocate the NH2 terminus across the membrane while type II signal-anchor proteins translocate the COOH terminus. We investigated the requirements for cytosolic protein components and nucleotides for the membrane targeting and insertion of single-spanning type I signal-anchor proteins. Besides the ribosome, signal recognition particle (SRP), GTP, and rough microsomes (RMs) no other components were found to be required. The GTP analogue GMPPNP could substitute for GTP in supporting the membrane insertion of IMC-CAT. By using a photocrosslinking assay we show that for secreted, type I and type II signal-anchor proteins the presence of both GTP and RMs is required for the release of the nascent chain from the 54-kD subunit of SRP. For two of the proteins studied the release of the nascent chain from SRP54 was accompanied by a new interaction with components of the ER. We conclude that the GTP-dependent release of the nascent chain from SRP54 occurs in an identical manner for each of the proteins studied.

MeSH Terms
Animals Apyrase/pharmacology Base Sequence Biological Transport Cell-Free System Endoplasmic Reticulum/metabolism Guanosine Triphosphate/physiology Guanylyl Imidodiphosphate/metabolism In Vitro Techniques Membrane Proteins/metabolism Molecular Sequence Data Oligonucleotides/chemistry Protein Precursors/metabolism Protein Sorting Signals/physiology Receptors, Transferrin/metabolism Recombinant Proteins/metabolism Ribonucleoproteins/metabolism Ribosomes/metabolism Signal Recognition Particle Structure-Activity Relationship
Chemicals
Membrane Proteins Oligonucleotides Protein Precursors Protein Sorting Signals Receptors, Transferrin Recombinant Proteins Ribonucleoproteins Signal Recognition Particle Guanylyl Imidodiphosphate Guanosine Triphosphate Apyrase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
High S
European Molecular Biology Laboratory, Heidelberg, Germany.
Flint N
Dobberstein B
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.113.1.25
Published
1991-04-00
Pages
25-34
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2288911
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.113.1.25 PMC Full Text View journal details
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