Publication — IRIC

Affinity purification of protein complexes from Drosophila embryos in cell cycle studies.

The ability to identify protein interactions is key to elucidating the molecular mechanisms of cellular processes, including mitosis and cell cycle regulation. Drosophila melanogaster, as a model system, provides powerful tools to study cell division using genetics, microscopy, and RNAi. Drosophila early embryos are highly enriched in mitotic protein complexes as their nuclei undergo 13 rounds of rapid, synchronous mitotic nuclear divisions in a syncytium during the first 2 h of development. Here, we describe simple methods for the affinity purification of protein complexes from transgenic fly embryos via protein A- and green fluorescent protein-tags fused to bait proteins of interest. This in vivo proteomics approach has allowed the identification of several known and novel mitotic protein interactions using mass spectrometry, and it expands the use of the Drosophila model in modern molecular biology.

Date de publication
14 mai 2014
Chercheurs
Lipinszki Z, Wang P, Grant R, Lindon C, Dzhindzhev NS, D'Avino PP, Przewloka MR, Glover DM, Archambault V
Référence PubMed
Methods Mol. Biol. 2014;1170:571-88
ID PubMed
24906338
Affiliation
Department of Genetics, University of Cambridge, Cambridge, UK.