Proteomics

Dataset Information

0

SILAC APMS analysis of RPAP3 C-term interactome

ABSTRACT: To get insights into the function of RPAP3, we characterized its partners by performing a proteomic analysis in human cells. We fused the Cterm part of RPAP3 domain to GFP and stably expressed it in HeLa cells using site-specific integration with the Flp-In system. Following differential labeling of GFP-RPAP3-Cter and control cells with isotopically labelled amino-acids (SILAC), whole cell extracts were immuno-precipitated (IP) with anti-GFP antibodies and immunoprecipitates were subjected to quantitative mass-spectrometry analysis. Similar experiments were then performed on two mutated form of RPAP3 that can no longer with the two essential AAA+ ATPases RUVBL1/RUVBL2, important partners of Wild type RPAP3 Cterm. 3 batchs of SILAC analysis K0R0 control Hela H9 vs K4R6 x-FLAG- GFP- RPAP3 Cterm Wild type K0R0 control Hela H9 vs K4R6 x-FLAG- GFP- RPAP3 Cterm mutant 1 R623A-M626A K0R0 control Hela H9 vs K4R6 x-FLAG- GFP- RPAP3 Cterm mutant 2 F630A-S632A

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

SUBMITTER: Franck Vandermoere  

LAB HEAD: Edouard Bertrand

PROVIDER: PXD009518 | Pride | 2018-06-10

REPOSITORIES: Pride

Json Xml

Dataset's files

Source:
Action DRS
Mut1_Search.tar Other
Mut1_andromeda.tar Other
Mut2_search.tar Other
PHAXRNASE_160223_1_E160315_01.raw Raw
PHAXRNASE_160223_1_E160315_02.raw Raw
Items per page:
1 - 5 of 39
altmetric image

Publications

R2TP is an HSP90 co-chaperone that assembles important macro-molecular machineries. It is composed of an RPAP3-PIH1D1 heterodimer, which binds the two essential AAA+ATPases RUVBL1/RUVBL2. Here, we resolve the structure of the conserved C-terminal domain of RPAP3, and we show that it directly binds RUVBL1/RUVBL2 hexamers. The human genome encodes two other proteins bearing RPAP3-C-terminal-like domains and three containing PIH-like domains. Systematic interaction analyses show that one RPAP3-like  ...[more]