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Interaction protoemics of Dictyostelium discoideum GFP-SAC1

ABSTRACT: Label free interaction protoemics of Dictyostelium discoideum GFP-SAC1 compared to GFP expressing cells

INSTRUMENT(S): Q Exactive

ORGANISM(S): Dictyostelium Discoideum (slime Mold)

SUBMITTER: Florian Fröhlich

LAB HEAD: Florian Fröhlich

PROVIDER: PXD039848 | Pride | 2023-12-01

REPOSITORIES: Pride

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			Action	DRS
	Control-a.raw	Raw
	Control-b.raw	Raw
	Control-c.raw	Raw
	Sac1-a.raw	Raw
	Sac1-b.raw	Raw

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Publications

Structure of the ceramide-bound SPOTS complex.

Schäfer Jan-Hannes JH Körner Carolin C Esch Bianca M BM Limar Sergej S Parey Kristian K Walter Stefan S Januliene Dovile D Moeller Arne A Fröhlich Florian F

Nature communications 20231004 1

Sphingolipids are structural membrane components that also function in cellular stress responses. The serine palmitoyltransferase (SPT) catalyzes the rate-limiting step in sphingolipid biogenesis. Its activity is tightly regulated through multiple binding partners, including Tsc3, Orm proteins, ceramides, and the phosphatidylinositol-4-phosphate (PI4P) phosphatase Sac1. The structural organization and regulatory mechanisms of this complex are not yet understood. Here, we report the high-resoluti ...[more]

PMID: 37794019

Similar Datasets

Identification of distinct active pools of yeast serine palmitoyltransferase in sub-compartments of the ER.

Project description:Interaction proteomics of rewired serine palmitoyl transferase strains from Esch et al. 2023

2023-12-01 | PXD047438 | Pride

Sac1 interaction proteomics of ALFA Sac1 and mutants

Project description:Sphingolipids are structural membrane components that also function in cellular stress responses. The serine palmitoyl-transferase (SPT) catalyzes the rate limiting step in sphingolipid biogenesis. Its activity is tightly regulated through multiple bind-ing partners, including Tsc3, Orm proteins, ceramides, and the phosphatidylinositol-4-phosphate (PI4P) phosphatase Sac1. The structural organization and regulatory mechanisms of this complex are not yet understood. Here, we report the high-resolution cryo-EM structures of the yeast SPT in complex with Tsc3 and Orm1 (SPOT) as dimers and monomers and a monomeric complex further carrying Sac1 (SPOTS). In all complexes, the tight interaction of the down-stream metabolite ceramide and Orm1 reveals the ceramide dependent inhibition. Additionally, observation of ceramide and ergosterol binding suggests a co-regulation of sphingolipid biogenesis and sterol metabolism within the SPOTS com-plex.

2023-10-24 | PXD044586 | Pride

Svf1-GFP pulldowns from Limar et al. 2023 JCB

Project description:Svf1-GFP pulldowns compared to untegged WT cells from SILAC labelled yeast cells. See Limar et al. 2023 JCB

2023-03-06 | PXD039395 | Pride

Yeast Serine palmitoyl transferase interaction proteomics

Project description:ALFA pulldowns of the Lcb1 subunit of the yeast serine palmitoyltransferase. Wt Lcb1 is compared to a mutant lacking the trans membrane helix 1 of Lcb1 and to control pulldowns lackingthe ALFA tag on Lcb1.

2024-09-12 | PXD048754 | Pride

Characterization of RBM10 interactome in unchallenged and MK1775-treated cells

Project description:Mapping of RBM10-interactome using GFP-trap purification of EGFP-RBM10 expressed in HEK293T cells before and after WEE1 inhibition by MK1775. HEK293T expressing EGFP only or EGFP-RBM10 fusion (2 main isoforms) were left untreated or treated with 300nM MK1775 for 4 hours prior to GFP-trap enrichment and mass spectrometry analysis.

2024-07-16 | PXD040109 | Pride

Lable-free LC-MSMS in HeLa on DYRK3 and Sec16A, Human cervical cancer cells

Project description:In order to identify sites of the SEC16A specifically phosphorylated by the Dual Specificity Kinase DYRK3, mCherry-SEC16A was transiently over expressed in HeLa cells together with GFP-DYRK3 or EGFP-C1 only

2023-06-23 | PXD041620 | Pride

Interaction proteomics of yeast Sac1-ALFA

Project description:Interaction proteomics of yeast Sac1-ALFA compared to untagged control strains

2023-12-01 | PXD039849 | Pride

phospho-proteomic data on the LATS1 interactome from cells treated with two proapoptotic signals: FAS and etoposide

Project description:Using this approach, we obtained phospho-proteomic data on the LATS1 interactome from cells treated with two proapoptotic signals: FAS and etoposide, which both activate LATS kinase activity [28].To identify the LATS1 interactome we transiently expressed GFP-LATS1 in HeLa cells, immunoprecipitated GFP-LATS1 with anti-GFP antibodies and identified the associated proteins using mass-spectrometry. Unspecific binding proteins were discarded by comparing with the control GFP IP.

2021-09-09 | PXD018903 | Pride

SILAC APMS analysis of RPAP3 C-term interactome

Project description: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

2018-06-10 | PXD009518 | Pride

SILAC APMS analysis of DNAAF2 interactome

Project description:To get insights into the function of DNAAF2 protein, we characterized its partners by performing a proteomic analysis in human cells. We fused this domain to GFP and stably expressed it in HeLa cells using site-specific integration with the Flp-In system. Following differential labeling of GFP- DNAAF2 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. SILAC labels: K0R0 control Hela H9 vs K4R6 x-FLAG- GFP- DNAAF2

2018-06-10 | PXD009501 | Pride

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