Biochemical isolation of Argonaute protein complexes by Ago-APP.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26351695.
- Also identified by DOI 10.1073/pnas.1506116112 and PMC identifier 4586862.
- No licence information is recorded for this record.
- Because redistribution is not established, this page shows the abstract only. Follow the links below for the full text.
Abstract
During microRNA (miRNA)-guided gene silencing, Argonaute (Ago) proteins interact with a member of the TNRC6/GW protein family. Here we used a short GW protein-derived peptide fused to GST and demonstrate that it binds to Ago proteins with high affinity. This allows for the simultaneous isolation of all Ago protein complexes expressed in diverse species to identify associated proteins, small RNAs, or target mRNAs. We refer to our method as "Ago protein Affinity Purification by Peptides" (Ago-APP). Furthermore, expression of this peptide competes for endogenous TNRC6 proteins, leading to global inhibition of miRNA function in mammalian cells.
Medical subject headings
- Argonaute Proteins
- Chromatography, Affinity
- Multiprotein Complexes
- Peptides