English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Selective cross-linking of interacting proteins using self-labeling tags.

Gautier, A., Nakata, E., Lukinavicius, G., Tan, K. T., & Johnsson, K. (2009). Selective cross-linking of interacting proteins using self-labeling tags. Journal of the American Chemical Society, 131(49), 17954-17962. doi:10.1021/ja907818q.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/21.11116/0000-0001-DFDA-F Version Permalink: http://hdl.handle.net/21.11116/0000-0001-DFE7-0
Genre: Journal Article

Files

show Files
hide Files
:
2629774.pdf (Publisher version), 3MB
 
File Permalink:
-
Name:
2629774.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute), Göttingen; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
2629774_Suppl.pdf (Supplementary material), 3MB
Name:
2629774_Suppl.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Gautier, A., Author
Nakata, E., Author
Lukinavicius, G.1, Author              
Tan, K. T., Author
Johnsson, K., Author
Affiliations:
1Laboratory of Chromatin Labeling and Imaging, Max Planck Institute for Biophysical Chemistry, Max Planck Society, ou_2616691              

Content

show
hide
Free keywords: -
 Abstract: We have designed molecules that permit the selective cross-linking (S-CROSS) of interacting proteins in cell lysates and the sensitive detection of the trapped complexes through in-gel fluorescence scanning. S-CROSS requires the expression of the putative interacting proteins as fusion to CLIP-tag or SNAP-tag, two protein tags that can be specifically labeled with synthetic probes. Bifunctional molecules that contain the substrates of the two tags connected via a fluorophore are used to selectively cross-link interacting proteins in cell lysate. The amount of trapped complex can be then quantified after SDS gel electrophoresis by in-gel fluorescence scanning. On the basis of a detailed kinetic analysis of the cross-linking reaction, we showed that the cross-linking efficiency can be used as an indicator of interaction between two proteins, allowing thereby the unambiguous identification of interacting protein pairs. We validated our approach by confirming a number of interactions through selective cross-linking and showed that it permits the quantitative and simultaneous analysis of multiple homotypic and heterotypic protein complexes and the differentiation between strong and weak protein−protein interactions.

Details

show
hide
Language(s): eng - English
 Dates: 2009-11-162009
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1021/ja907818q
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of the American Chemical Society
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 131 (49) Sequence Number: - Start / End Page: 17954 - 17962 Identifier: -