Boosting tandem affinity purification of plant protein complexes

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Affinity purification of protein complexes.

INTRODUCTION Knowledge of the composition of protein complexes provides key insights into their functions. Immunoaffinity purification provides an effective means for isolating protein complexes and elucidating their composition. Immunoisolation is achieved with antibodies directed either specifically against the proteins of interest or against tags that are coupled to the proteins of interest....

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We employed a combination of tandem affinity purification and mass spectrometry for deciphering protein complexes and the protein interaction network in budding yeast. 53 genes were epitope-tagged, and their interaction partners were isolated by two-step immunoaffinity chromatography from whole cell lysates. 38 baits pulled down a total of 220 interaction partners, which are members of 19 funct...

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Tandem affinity purification combined with mass spectrometry to identify components of protein complexes.

Most biological processes are governed by multiprotein complexes rather than individual proteins. Identification of protein complexes therefore is becoming increasingly important to gain a molecular understanding of cells and organisms. Mass spectrometry-based proteomics combined with affinity-tag-based protein purification is one of the most effective strategies to isolate and identify protein...

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Discovery of Protein Complexes with Core-Attachment Structures from Tandem Affinity Purification (TAP) Data

Many cellular functions involve protein complexes that are formed by multiple interacting proteins. Tandem Affinity Purification (TAP) is a popular experimental method for detecting such multi-protein interactions. However, current computational methods that predict protein complexes from TAP data require converting the co-complex relationships in TAP data into binary interactions. The resultin...

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Interaction proteomics: characterization of protein complexes using tandem affinity purification-mass spectrometry.

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ژورنال

عنوان ژورنال: Trends in Plant Science

سال: 2008

ISSN: 1360-1385

DOI: 10.1016/j.tplants.2008.08.002