Dataset Information

Quantitative GTPase affinity purification identifies Rho family protein interaction partners

ABSTRACT: Although Rho GTPases are essential molecular switches involved in many cellular processes, an unbiased experimental comparison of their interaction partners was not yet performed. Here, we develop quantitative GTPase affinity purification (qGAP) to systematically identify interaction partners of six Rho GTPases (Cdc42, Rac1, RhoA, RhoB, RhoC, RhoD) depending on their nucleotide loading state. We use Stable isotope labeling by Amino acids in cell culture (SILAC) and label free quantification (LFQ). Our interaction network contains many new proteins, reveals highly promiscuous binding of several effectors and mirrors evolutionary relationships of Rho GTPases.

INSTRUMENT(S): LTQ Orbitrap, Q Exactive

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Brain, Cell Culture

SUBMITTER: Florian Paul

LAB HEAD: Dr Matthias Selbach

PROVIDER: PXD004190 | Pride | 2016-12-23

REPOSITORIES: pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20100307_FEP_89_NHS_PD100305_proteinGroups_Analyse1305_SILAC.zip	Other
	201108_brainlysates_RhoARac1Cdc42_1305_LFQ1.zip	Other
	Bert_20100307_FEP_89_NHS_PD100305_RhoA_ST.RAW	Raw
	Bert_20100307_FEP_90_NHS_PD100305_RhoA_CR.RAW	Raw
	Bert_20100307_FEP_91_NHS_PD100305_Rac1_ST.RAW	Raw

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Publications

Quantitative GTPase Affinity Purification Identifies Rho Family Protein Interaction Partners.

Paul Florian F Zauber Henrik H von Berg Laura L Rocks Oliver O Daumke Oliver O Selbach Matthias M

Molecular & cellular proteomics : MCP 20161116 1

Although Rho GTPases are essential molecular switches involved in many cellular processes, an unbiased experimental comparison of their interaction partners was not yet performed. Here, we develop quantitative GTPase affinity purification (qGAP) to systematically identify interaction partners of six Rho GTPases (Cdc42, Rac1, RhoA, RhoB, RhoC, and RhoD), depending on their nucleotide loading state. The method works with cell line or tissue-derived protein lysates in combination with SILAC-based o ...[more]

PMID: 27852748

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