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

Affinity purification of protein complexes from Drosophila embryos in cell cycle studies
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.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.
1064-3745
571-588
Springer
Lipinszki, Zoltan
e405fd6d-1bee-4725-96b7-7c315f2826ed
Wang, Peng
be7926cf-52df-44e9-b122-f37812673b38
Grant, Rhys
41bcb086-c7c3-411b-92e1-2ff643de4f69
Lindon, Catherine
6d9022fd-b2b7-4486-91d2-aaa0409dbce0
Dzhindzhev, Nikola S.
9e198725-621e-421e-bed0-0b654ca94dd8
D'Avino, Pier Paolo
0e659e0c-c3a7-4344-8850-3364810b9445
Przewloka, Marcin
9b25e73c-ec15-43df-a5a4-ac9574bb20ab
Glover, David M.
cca9cd19-3e1e-4906-b418-41876c1f9c61
Archambault, Vincent
3654cc6d-d3f9-4404-8ae5-9d8af8b008dc
Noguchi, Eishi
Gadaleta, Mariana C.
Lipinszki, Zoltan
e405fd6d-1bee-4725-96b7-7c315f2826ed
Wang, Peng
be7926cf-52df-44e9-b122-f37812673b38
Grant, Rhys
41bcb086-c7c3-411b-92e1-2ff643de4f69
Lindon, Catherine
6d9022fd-b2b7-4486-91d2-aaa0409dbce0
Dzhindzhev, Nikola S.
9e198725-621e-421e-bed0-0b654ca94dd8
D'Avino, Pier Paolo
0e659e0c-c3a7-4344-8850-3364810b9445
Przewloka, Marcin
9b25e73c-ec15-43df-a5a4-ac9574bb20ab
Glover, David M.
cca9cd19-3e1e-4906-b418-41876c1f9c61
Archambault, Vincent
3654cc6d-d3f9-4404-8ae5-9d8af8b008dc
Noguchi, Eishi
Gadaleta, Mariana C.

Lipinszki, Zoltan, Wang, Peng, Grant, Rhys, Lindon, Catherine, Dzhindzhev, Nikola S., D'Avino, Pier Paolo, Przewloka, Marcin, Glover, David M. and Archambault, Vincent (2014) Affinity purification of protein complexes from Drosophila embryos in cell cycle studies. In, Noguchi, Eishi and Gadaleta, Mariana C. (eds.) Cell Cycle Control. (Methods in Molecular Biology, , (doi:10.1007/978-1-4939-0888-2_33), 1170) New York, US. Springer, pp. 571-588. (doi:10.1007/978-1-4939-0888-2_33).

Record type: Book Section

Abstract

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.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.

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Published date: 2014
Organisations: Molecular and Cellular, Centre for Biological Sciences

Identifiers

Local EPrints ID: 393859
URI: http://eprints.soton.ac.uk/id/eprint/393859
ISSN: 1064-3745
PURE UUID: 4db030e4-1fb1-4782-b9bc-cbb83d1d1c88
ORCID for Marcin Przewloka: ORCID iD orcid.org/0000-0002-0329-9162

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Date deposited: 09 May 2016 09:31
Last modified: 20 Jul 2019 00:33

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Contributors

Author: Zoltan Lipinszki
Author: Peng Wang
Author: Rhys Grant
Author: Catherine Lindon
Author: Nikola S. Dzhindzhev
Author: Pier Paolo D'Avino
Author: David M. Glover
Author: Vincent Archambault
Editor: Eishi Noguchi
Editor: Mariana C. Gadaleta

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