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Identifying RasGAP SH2 binding partners in Drosophila using mass spectrometry

ABSTRACT: Drosophila S2 cells were transfected with RasGAP(WT), RasGAP(SH2*32*) and GFP control. 4 mg of S2 cell lysate was used to pulldown LAP and GFP tagged constructs using GFP-Trap. The pulled down fraction was loaded on SDS-PAGE gels and coomassie stained before several bands (a-u) from two independent but parallel experiments were excised and subjected to MS analysis.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

SUBMITTER: Behzad Rowshanravan

LAB HEAD: Behzad Rowshanravan

PROVIDER: PXD000176 | Pride | 2014-05-20

REPOSITORIES: Pride

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			Action	DRS
	PRIDE_Exp_Complete_Ac_28714.pride.mgf.gz	Mgf
	PRIDE_Exp_Complete_Ac_28714.pride.mztab.gz	Mztab
	PRIDE_Exp_Complete_Ac_28714.xml.gz	Xml
	PRIDE_Exp_Complete_Ac_28715.pride.mgf.gz	Mgf
	PRIDE_Exp_Complete_Ac_28715.pride.mztab.gz	Mztab

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Publications

RasGAP mediates neuronal survival in Drosophila through direct regulation of Rab5-dependent endocytosis.

Rowshanravan Behzad B Woodcock Simon A SA Botella José A JA Kiermayer Claudia C Schneuwly Stephan S Hughes David A DA

Journal of cell science 20140509 Pt 13

The GTPase Ras can either promote or inhibit cell survival. Inactivating mutations in Drosophila RasGAP (encoded by vap), a Ras GTPase-activating protein, lead to age-related brain degeneration. Genetic interactions implicate the epidermal growth factor receptor (EGFR)-Ras pathway in promoting neurodegeneration but the mechanism is not known. Here, we show that the Src homology 2 (SH2) domains of RasGAP are essential for its neuroprotective function. By using affinity purification and mass spect ...[more]

PMID: 24816559

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