English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Systematic Protein-Protein Interaction Analysis Reveals Intersubcomplex Contacts in the Nuclear Pore Complex

Apelt, L., Knockenhauer, K. E., Leksa, N. C., Benlasfer, N., Schwartz, T. U., & Stelzl, U. (2016). Systematic Protein-Protein Interaction Analysis Reveals Intersubcomplex Contacts in the Nuclear Pore Complex. Molecular and Cellular Proteomics, 15(8), 2594-2606. doi:10.1074/mcp.M115.054627.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002B-4ADC-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002B-4ADD-8
Genre: Journal Article

Files

show Files
hide Files
:
Apelt.pdf (Publisher version), 3MB
Name:
Apelt.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
© 2016 American Society for Biochemistry and Molecular Biology
License:
-

Locators

show

Creators

show
hide
 Creators:
Apelt, Luise1, Author              
Knockenhauer, Kevin E. , Author
Leksa, Nina C. , Author
Benlasfer, Nouhad1, Author              
Schwartz, Thomas U. , Author
Stelzl, Ulrich1, 2, Author              
Affiliations:
1Molecular Interaction Networks (Ulrich Stelzl), Independent Junior Research Groups (OWL), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479660              
2Institute of Pharmaceutical Sciences, Pharmaceutical Chemistry, University of Graz, Graz, Austria, ou_persistent22              

Content

show
hide
Free keywords: -
 Abstract: The nuclear pore complex (NPC) enables transport across the nuclear envelope. It is one of the largest multiprotein assemblies in the cell, built from about 30 proteins called nucleoporins (Nups), organized into distinct subcomplexes. Structure determination of the NPC is a major research goal. The assembled ∼40-112 MDa NPC can be visualized by cryoelectron tomography (cryo-ET), while Nup subcomplexes are studied crystallographically. Docking the crystal structures into the cryo-ET maps is difficult because of limited resolution. Further, intersubcomplex contacts are not well characterized. Here, we systematically investigated direct interactions between Nups. In a comprehensive, structure-based, yeast two-hybrid interaction matrix screen, we mapped protein-protein interactions in yeast and human. Benchmarking against crystallographic and coaffinity purification data from the literature demonstrated the high coverage and accuracy of the data set. Novel intersubcomplex interactions were validated biophysically in microscale thermophoresis experiments and in intact cells through protein fragment complementation. These intersubcomplex interaction data provide direct experimental evidence toward possible structural arrangements of architectural elements within the assembled NPC, or they may point to assembly intermediates. Our data favors an assembly model in which major architectural elements of the NPC, notably the Y-complex, exist in different structural contexts within the scaffold.

Details

show
hide
Language(s): eng - English
 Dates: 2016-05-182016-08
 Publication Status: Published in print
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: DOI: 10.1074/mcp.M115.054627
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Molecular and Cellular Proteomics
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Bethesda, MD : American Society for Biochemistry and Molecular Biology
Pages: - Volume / Issue: 15 (8) Sequence Number: - Start / End Page: 2594 - 2606 Identifier: ISSN: 1535-9476
CoNE: /journals/resource/111035577487002