Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin β1
Joint Authors
Kast, Juergen
Klockenbusch, Cordula
Source
Issue
Vol. 2010, Issue 2010 (31 Dec. 2009), pp.1-13, 13 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-04-29
Country of Publication
Egypt
No. of Pages
13
Main Subjects
Abstract EN
Formaldehyde cross-linking of protein complexes combined with immunoprecipitation and mass spectrometry analysis is a promising technique for analysing protein-protein interactions, including those of transient nature.
Here we used integrin β1 as a model to describe the application of formaldehyde cross-linking in detail, particularly focusing on the optimal parameters for cross-linking, the detection of formaldehyde cross-linked complexes, the utility of antibodies, and the identification of binding partners.
Integrin β1 was found in a high molecular weight complex after formaldehyde cross-linking.
Eight different anti-integrin β1 antibodies were used for pull-down experiments and no loss in precipitation efficiency after cross-linking was observed.
However, two of the antibodies could not precipitate the complex, probably due to hidden epitopes.
Formaldehyde cross-linked complexes, precipitated from Jurkat cells or human platelets and analyzed by mass spectrometry, were found to be composed of integrin β1, α4 and α6 or β1, α6, α2, and α5, respectively.
American Psychological Association (APA)
Klockenbusch, Cordula& Kast, Juergen. 2010. Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin β1. BioMed Research International،Vol. 2010, no. 2010, pp.1-13.
https://search.emarefa.net/detail/BIM-989188
Modern Language Association (MLA)
Klockenbusch, Cordula& Kast, Juergen. Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin β1. BioMed Research International No. 2010 (Dec. 2010), pp.1-13.
https://search.emarefa.net/detail/BIM-989188
American Medical Association (AMA)
Klockenbusch, Cordula& Kast, Juergen. Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin β1. BioMed Research International. 2010. Vol. 2010, no. 2010, pp.1-13.
https://search.emarefa.net/detail/BIM-989188
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-989188