SILAC-based proteomic quantification of chemoattractant-induced cytoskeleton dynamics on a second to minute timescale.

SILAC-based proteomic quantification of chemoattractant-induced cytoskeleton dynamics on a second to minute timescale.
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DOI:
10.1038/ncomms4319
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发表时间:
2014-02-26
影响因子:
16.6
通讯作者:
Weijer, Cornelis J.
Weijer, Cornelis J.
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Sobczyk, Grzegorz J.;Wang, Jun;Weijer, Cornelis J.

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Cytoskeletal dynamics during cell behaviours ranging from endocytosis and exocytosis to cell division and movement is controlled by a complex network of signalling pathways, the full details of which are as yet unresolved. Here we show that SILAC-based proteomic methods can be used to characterize the rapid chemoattractant-induced dynamic changes in the actin–myosin cytoskeleton and regulatory elements on a proteome-wide scale with a second to minute timescale resolution. This approach provides novel insights in the ensemble kinetics of key cytoskeletal constituents and association of known and novel identified binding proteins. We validate the proteomic data by detailed microscopy-based analysis of in vivo translocation dynamics for key signalling factors. This rapid large-scale proteomic approach may be applied to other situations where highly dynamic changes in complex cellular compartments are expected to play a key role. Actin-dependent motility is driven by the rapid changes in the recruitment of many different structural and regulatory proteins at the cell’s cortex. Sobczyk et al. characterize these changes in the cytoskeletal proteome on a second to minute timescale during chemotactic response in Dictyostelium using SILAC-based proteomics.
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