Cargando…

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...

Descripción completa

Detalles Bibliográficos
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1987
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114417/
https://www.ncbi.nlm.nih.gov/pubmed/3643215
_version_ 1782140415102681088
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