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Prostasomes - Proteomic profiling of detergent resistant membranes (lipid rafts) of prostasomes

ABSTRACT: Which proteins can be found in rafts enriched from human semen.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Bo Ek

LAB HEAD: Bo Ek

PROVIDER: PXD002163 | Pride | 2015-08-18

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	F015286.mzid.gz	Mzid
	F015286.pride.mztab.gz	Mztab
	data06.mgf	Mgf
	data06.pride.mgf.gz	Mgf
	data06.raw	Raw

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Publications

Proteomic Profiling of Detergent Resistant Membranes (Lipid Rafts) of Prostasomes.

Dubois Louise L Ronquist Karl K Göran KK Ek Bo B Ronquist Gunnar G Larsson Anders A

Molecular & cellular proteomics : MCP 20150813 11

Prostasomes are exosomes derived from prostate epithelial cells through exocytosis by multivesicular bodies. Prostasomes have a bilayered membrane and readily interact with sperm. The membrane lipid composition is unusual with a high contribution of sphingomyelin at the expense of phosphatidylcholine and saturated and monounsaturated fatty acids are dominant. Lipid rafts are liquid-ordered domains that are more tightly packed than the surrounding nonraft phase of the bilayer. Lipid rafts are pro ...[more]

PMID: 26272980

Similar Datasets

Proteomic Profiling of Detergent Resistant Membranes (Lipid Rafts) of Prostasomes.

Project description:Prostasomes are exosomes derived from prostate epithelial cells through exocytosis by multivesicular bodies. Prostasomes have a bilayered membrane and readily interact with sperm. The membrane lipid composition is unusual with a high contribution of sphingomyelin at the expense of phosphatidylcholine and saturated and monounsaturated fatty acids are dominant. Lipid rafts are liquid-ordered domains that are more tightly packed than the surrounding nonraft phase of the bilayer. Lipid rafts are proposed to be highly dynamic, submicroscopic assemblies that float freely within the liquid disordered membrane bilayer and some proteins preferentially partition into the ordered raft domains. We asked the question whether lipid rafts do exist in prostasomes and, if so, which proteins might be associated with them. Prostasomes of density range 1.13-1.19g/ml were subjected to density gradient ultracentrifugation in sucrose fabricated by phosphate buffered saline (PBS) containing 1% Triton X-100 with capacity for banding at 1.10 g/ml, i.e. the classical density of lipid rafts. Prepared prostasomal lipid rafts (by gradient ultracentrifugation) were analyzed by mass spectrometry. The clearly visible band on top of 1.10g/ml sucrose in the Triton X-100 containing gradient was subjected to liquid chromatography-tandem MS and more than 370 lipid raft associated proteins were identified. Several of them were involved in intraluminal vesicle formation, e.g. tetraspanins, ESCRTs, and Ras-related proteins. This is the first comprehensive liquid chromatography-tandem MS profiling of proteins in lipid rafts derived from exosomes. Data are available via ProteomeXchange with identifier PXD002163.

| S-EPMC4638043 | biostudies-literature

Proteomic Analysis of Lipid Raft-Like Detergent-Resistant Membranes of Lens Fiber Cells.

Project description:PurposePlasma membranes of lens fiber cells have high levels of long-chain saturated fatty acids, cholesterol, and sphingolipids-key components of lipid rafts. Thus, lipid rafts are expected to constitute a significant portion of fiber cell membranes and play important roles in lens biology. The purpose of this study was to characterize the lens lipid raft proteome.MethodsQuantitative proteomics, both label-free and iTRAQ methods, were used to characterize lens fiber cell lipid raft proteins. Detergent-resistant, lipid raft membrane (DRM) fractions were isolated by sucrose gradient centrifugation. To confirm protein localization to lipid rafts, protein sensitivity to cholesterol removal by methyl-?-cyclodextrin was quantified by iTRAQ analysis.ResultsA total of 506 proteins were identified in raft-like detergent-resistant membranes. Proteins identified support important functions of raft domains in fiber cells, including trafficking, signal transduction, and cytoskeletal organization. In cholesterol-sensitivity studies, 200 proteins were quantified and 71 proteins were strongly affected by cholesterol removal. Lipid raft markers flotillin-1 and flotillin-2 and a significant fraction of AQP0, MP20, and AQP5 were found in the DRM fraction and were highly sensitive to cholesterol removal. Connexins 46 and 50 were more abundant in nonraft fractions, but a small fraction of each was found in the DRM fraction and was strongly affected by cholesterol removal. Quantification of modified AQP0 confirmed that fatty acylation targeted this protein to membrane raft domains.ConclusionsThese data represent the first comprehensive profile of the lipid raft proteome of lens fiber cells and provide information on membrane protein organization in these cells.

| S-EPMC4699431 | biostudies-literature

Proteomic analysis of Escherichia coli detergent-resistant membranes (DRM).

Project description:Membrane microdomains or lipid rafts compartmentalize cellular processes by laterally organizing membrane components. Such sub-membrane structures were mainly described in eukaryotic cells, but, recently, also in bacteria. Here, the protein content of lipid rafts in Escherichia coli was explored by mass spectrometry analyses of Detergent Resistant Membranes (DRM). We report that at least three of the four E. coli flotillin homologous proteins were found to reside in DRM, along with 77 more proteins. Moreover, the proteomic data were validated by subcellular localization, using immunoblot assays and fluorescence microscopy of selected proteins. Our results confirm the existence of lipid raft-like microdomains in the inner membrane of E. coli and represent the first comprehensive profiling of proteins in these bacterial membrane platforms.

| S-EPMC6788730 | biostudies-literature

Caveolin-1 Endows Order in Cholesterol-Rich Detergent Resistant Membranes.

Project description:Cholesterol-enriched functional portions of plasma membranes, such as caveolae and rafts, were isolated from lungs of wild-type (WT) and caveolin-1 knockout (Cav-1 KO) mice within detergent resistant membranes (DRMs). To gain insight into their molecular composition we performed proteomic and lipid analysis on WT and Cav-1 KO-DRMs that showed predicted variations of proteomic profiles and negligible differences in lipid composition, while Langmuir monolayer technique and small and wide-angle X-ray scattering (SAXS-WAXS) were here originally introduced to study DRMs biophysical association state. Langmuir analysis of Cav-1 containing DRMs displayed an isotherm with a clear-cut feature, suggesting the coexistence of the liquid-ordered (Lo) phase typical of the raft structure, namely "cholesterol-rich Lo phase," with a phase fully missing in Cav-1 KO that we named "caveolin-induced Lo phase." Furthermore, while the sole lipid component of both WT and KO-DRMs showed qualitatively similar isotherm configuration, the reinsertion of recombinant Cav-1 into WT-DRMs lipids restored the WT-DRM pattern. X-ray diffraction results confirmed that Cav-1 causes the formation of a "caveolin-induced Lo phase," as suggested by Langmuir experiments, allowing us to speculate about a possible structural model. These results show that the unique molecular link between Cav-1 and cholesterol can spur functional order in a lipid bilayer strictly derived from biological sources.

| S-EPMC6680987 | biostudies-literature

Human platelet detergent resistant membranes proteome

Project description:Here we characterized the proteins associated with detergent resistant membranes (DRMs) from resting and activated human platelets. We achieved a proteomic profile of 141 DRM proteins involved in many biological pathways including 'PAR-1-mediated thrombin signal events', 'Integrin family cell surface interactions', 'Platelet activation, signalling and aggregation' and 'Platelet degranulation'. Therefore, a high level of relevant platelet signalling and trafficking proteins were identified e.g., Rap 1b, Src, SNAP-23 and syntaxin-11, implicating platelet DRMs as concentrating platforms not only in the critical platelet function of activation but also in secretion/degranulation.

2017-03-28 | MSV000080708 | MassIVE

Prostasomes

Project description:Which proteins can be found in rafts enriched from human semen.

2016-07-19 | MSV000079918 | MassIVE

Human platelet detergent resistant membranes proteome

2016-06-23 | PXD002554 | Pride

Detergent-resistant membranes in Agrobacterium tumefaciens

Project description:We established a protocol for the biochemical fractionation of Agrobacterium tumefaciens membranes, and characterized the proteome of detergent-sensitive and detergent-resistant membrane fractions by mass spectrometry. We found that well-established microdomain marker proteins co-purify with proteins associated with transport or secretion processes in the detergent-resistant fraction, suggesting a functional micro-compartmentalization of the Gram-negative membrane. The presence of components of virulence-associated secretion systems in detergent-resistant membranes supports the involvement of bacterial microdomains in host-microbe interaction and microbial competition.

2021-11-03 | PXD028782 | Pride

Reversible Dissolution of Microdomains in Detergent-Resistant Membranes at Physiological Temperature.

Project description:The formation of lipid microdomains ("rafts") is presumed to play an important role in various cellular functions, but their nature remains controversial. Here we report on microdomain formation in isolated, detergent-resistant membranes from MDA-MB-231 human breast cancer cells, studied by atomic force microscopy (AFM). Whereas microdomains were readily observed at room temperature, they shrunk in size and mostly disappeared at higher temperatures. This shrinking in microdomain size was accompanied by a gradual reduction of the height difference between the microdomains and the surrounding membrane, consistent with the behaviour expected for lipids that are laterally segregated in liquid ordered and liquid disordered domains. Immunolabeling experiments demonstrated that the microdomains contained flotillin-1, a protein associated with lipid rafts. The microdomains reversibly dissolved and reappeared, respectively, on heating to and cooling below temperatures around 37 °C, which is indicative of radical changes in local membrane order close to physiological temperature.

| S-EPMC4493071 | biostudies-literature

Identification of novel ?-secretase-associated proteins in detergent-resistant membranes from brain.

Project description:In Alzheimer disease, oligomeric amyloid ?-peptide (A?) species lead to synapse loss and neuronal death. ?-Secretase, the transmembrane protease complex that mediates the final catalytic step that liberates A? from its precursor protein (APP), has a multitude of substrates, and therapeutics aimed at reducing A? production should ideally be specific for APP cleavage. It has been shown that APP can be processed in lipid rafts, and ?-secretase-associated proteins can affect A? production. Here, we use a biotinylated inhibitor for affinity purification of ?-secretase and associated proteins and mass spectrometry for identification of the purified proteins, and we identify novel ?-secretase-associated proteins in detergent-resistant membranes from brain. Furthermore, we show by small interfering RNA-mediated knockdown of gene expression that a subset of the ?-secretase-associated proteins, in particular voltage-dependent anion channel 1 (VDAC1) and contactin-associated protein 1 (CNTNAP1), reduced A? production (A?40 and A?42) by around 70%, whereas knockdown of presenilin 1, one of the essential ?-secretase complex components, reduced A? production by 50%. Importantly, these proteins had a less pronounced effect on Notch processing. We conclude that VDAC1 and CNTNAP1 associate with ?-secretase in detergent-resistant membranes and affect APP processing and suggest that molecules that interfere with this interaction could be of therapeutic use for Alzheimer disease.

| S-EPMC3320946 | biostudies-literature

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