ArticleJournal of biomedicine & biotechnology2010
Optimization of formaldehyde cross-linking for protein interaction analysis of non-tagged integrin beta1.
Article in Journal of biomedicine & biotechnology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 75 papers.
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Who cites it
75 citing papers in PubMed, 144 citations in OpenAlex.
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- Altered sulfation status of FAM20C-dependent chondroitin sulfate is associated with osteosclerotic bone dysplasia.Nature communications · 2022Article
- ESRP1-regulated isoform switching of LRRFIP2 determines metastasis of gastric cancer.Nature communications · 2022Article
- Label-Free Quantification of Molecular Interaction in Live Red Blood Cells by Tracking Nanometer Scale Membrane Fluctuations.Small (Weinheim an der Bergstrasse, Germany) · 2022Article
- Biomimetic mimicry of formaldehyde-induced DNA-protein crosslinks in the confined space of a metal-organic framework.Chemical science · 2022Article
- VDAC1 regulates neuronal cell loss after retinal trauma injury by a mitochondria-independent pathway.Cell death & disease · 2022Article
- A balanced Oct4 interactome is crucial for maintaining pluripotency.Science advances · 2022Article
- In Vivo Protein Cross-Linking and Coimmunoprecipitation in Haloferax volcanii.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Comprehensive Atlas of the Myelin Basic Protein Interaction Landscape.Biomolecules · 2021Article
- Searching for Biological Function of the Mysterious PA2504 Protein fromInternational journal of molecular sciences · 2021Article
- New Insights on NETosis Induced byAntioxidants (Basel, Switzerland) · 2021Article
- Dbnl and β-catenin promote pro-N-cadherin processing to maintain apico-basal polarity.The Journal of cell biology · 2021Article
15 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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 beta1 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 beta1 was found in a high molecular weight complex after formaldehyde cross-linking. Eight different anti-integrin beta1 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 beta1, alpha4 and alpha6 or beta1, alpha6, alpha2, and alpha5, respectively.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.