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Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking
We have studied the interaction between the signal sequence of nascent preprolactin and the signal recognition particle (SRP) during the initial events in protein translocation across the endoplasmic reticulum membrane. A new method of affinity labeling was used, whereby lysine residues, carrying th...
Formato: | Texto |
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Lenguaje: | English |
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The Rockefeller University Press
1987
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114417/ https://www.ncbi.nlm.nih.gov/pubmed/3643215 |
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collection | PubMed |
description | We have studied the interaction between the signal sequence of nascent preprolactin and the signal recognition particle (SRP) during the initial events in protein translocation across the endoplasmic reticulum membrane. A new method of affinity labeling was used, whereby lysine residues, carrying the photoreactive group 4-(3- trifluoromethyldiazirino) benzoic acid in their side chains, are incorporated into a protein by means of modified lysyl-tRNA, and cross- linking to the interacting component is induced by irradiation. SRP interacts through its Mr 54,000 polypeptide component with the signal sequences of nascent preprolactin chains containing about 70 residues, and with decreasing affinity with longer chains as well; it causes inhibition of elongation. Binding of SRP is reversible and requires the nascent chain to be bound to a functional ribosome. SRP cross-linked to the signal sequence still inhibits elongation but does not prevent it completely. We conclude that SRP does not block the exit site of the polypeptide chain on the ribosome. The SRP receptor of the endoplasmic reticulum membrane displaces the signal sequence from SRP and, even if SRP is cross-linked, releases elongation arrest. |
format | Text |
id | pubmed-2114417 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1987 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21144172008-05-01 Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking J Cell Biol Articles We have studied the interaction between the signal sequence of nascent preprolactin and the signal recognition particle (SRP) during the initial events in protein translocation across the endoplasmic reticulum membrane. A new method of affinity labeling was used, whereby lysine residues, carrying the photoreactive group 4-(3- trifluoromethyldiazirino) benzoic acid in their side chains, are incorporated into a protein by means of modified lysyl-tRNA, and cross- linking to the interacting component is induced by irradiation. SRP interacts through its Mr 54,000 polypeptide component with the signal sequences of nascent preprolactin chains containing about 70 residues, and with decreasing affinity with longer chains as well; it causes inhibition of elongation. Binding of SRP is reversible and requires the nascent chain to be bound to a functional ribosome. SRP cross-linked to the signal sequence still inhibits elongation but does not prevent it completely. We conclude that SRP does not block the exit site of the polypeptide chain on the ribosome. The SRP receptor of the endoplasmic reticulum membrane displaces the signal sequence from SRP and, even if SRP is cross-linked, releases elongation arrest. The Rockefeller University Press 1987-02-01 /pmc/articles/PMC2114417/ /pubmed/3643215 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
title | Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
title_full | Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
title_fullStr | Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
title_full_unstemmed | Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
title_short | Direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
title_sort | direct probing of the interaction between the signal sequence of nascent preprolactin and the signal recognition particle by specific cross-linking |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114417/ https://www.ncbi.nlm.nih.gov/pubmed/3643215 |