ArticleJournal of visualized experiments : JoVE2013
Identifying protein-protein interaction in Drosophila adult heads by Tandem Affinity Purification (TAP).
Article in Journal of visualized experiments : JoVE, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
7 citing papers in PubMed, 9 citations in OpenAlex.
- An RNA-immunoprecipitation protocol to identify RNAs associated with RNA-binding proteins in cytoplasmic and nuclearSTAR protocols · 2022Article
- C9orf72 arginine-rich dipeptide proteins interact with ribosomal proteins in vivo to induce a toxic translational arrest that is rescued by eIF1A.Acta neuropathologica · 2019Article
- Mitochondrial dysfunction induces dendritic loss via eIF2α phosphorylation.The Journal of cell biology · 2017Article
- Arl2- and Msps-dependent microtubule growth governs asymmetric division.The Journal of cell biology · 2016Article
- Mask loss-of-function rescues mitochondrial impairment and muscle degeneration of Drosophila pink1 and parkin mutants.Human molecular genetics · 2015Article
- SkpA restrains synaptic terminal growth during development and promotes axonal degeneration following injury.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2014Article
- Drosophila Syd-1, liprin-α, and protein phosphatase 2A B' subunit Wrd function in a linear pathway to prevent ectopic accumulation of synaptic materials in distal axons.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2014Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Genetic screens conducted using Drosophila melanogaster (fruit fly) have made numerous milestone discoveries in the advance of biological sciences. However, the use of biochemical screens aimed at extending the knowledge gained from genetic analysis was explored only recently. Here we describe a method to purify the protein complex that associates with any protein of interest from adult fly heads. This method takes advantage of the Drosophila GAL4/UAS system to express a bait protein fused with a Tandem Affinity Purification (TAP) tag in fly neurons in vivo, and then implements two rounds of purification using a TAP procedure similar to the one originally established in yeast(1) to purify the interacting protein complex. At the end of this procedure, a mixture of multiple protein complexes is obtained whose molecular identities can be determined by mass spectrometry. Validation of the candidate proteins will benefit from the resource and ease of performing loss-of-function studies in flies. Similar approaches can be applied to other fly tissues. We believe that the combination of genetic manipulations and this proteomic approach in the fly model system holds tremendous potential for tackling fundamental problems in the field of neurobiology and beyond.
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Registered trials
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