Proteomic Analysis of Paired Human Milk Fat Globules and Milk Fat Globule Membranes.
Ontology highlight
ABSTRACT:
INSTRUMENT(S): timsTOF Pro 2
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: 
Jim McManaman
PROVIDER: MSV000092892 | MassIVE |
REPOSITORIES: MassIVE
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Project description:Intake of high-fat foods raises postprandial plasma triglycerides and inflammatory markers, which may depend on the type of fat ingested. Dairy products are commonly consumed, but not much is known about the impact of milk fat and the milk fat globule membrane on postprandial inflammation. Here, we aimed to study the effect of milk fat with and without milk fat globule membrane and vegetable blend fat on post-prandial inflammation, with a focus on blood monocyte gene expression. We performed a randomized, double-blind cross-over trial in middle-aged healthy male and female volunteers (BMI 22–27 kg/m2). The participants consumed a meal shake containing 95.5 g of fat consisting of either a vegetable fat blend (VEGE), anhydrous milk fat (AMF, without milk fat globule membrane), or cream (CREAM, containing milk fat globule membrane). Blood monocytes were collected at 0 and 6 hours postprandially and used for bulk RNA sequencing and ex vivo stimulation with LPS. Consumption of all three shakes significantly decreased the percentage of classical monocytes and increased the percentages of intermediate monocytes and non-classical monocytes. No differences in these measures were observed between shakes. Using a threshold of p < 0.01, 787 genes were differentially regulated postprandially between the three shakes. 89 genes were differentially regulated postprandially between AMF and VEGE, 373 genes between AMF and CREAM, and 667 genes between VEGE and CREAM, indicating that the effect of CREAM on monocyte gene expression was distinct from AMF and VEGE. Pathway analyses showed that VEGE significantly increased the expression of genes involved in inflammatory pathways, whereas this was less pronounced after AMF and not observed after CREAM. In addition, CREAM significantly down-regulated the expression of genes involved in energy metabolism-related pathways, such as glycolysis, TCA cycle, and oxidative phosphorylation, as well as HIF1 signaling. Compared to the consumption of an anhydrous milk fat without milk fat globule membrane and a vegetable fat blend, the consumption of cream with milk fat globule membrane downregulated inflammatory pathways in blood monocytes, thus suggesting a potential inflammation inhibitory effect of milk fat globule membrane.
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Project description:Camel milk is widely characterized with regard to caseins and whey proteins. However, in camelids, close to nothing is known about the Milk Fat Globule Membrane (MFGM), the membrane surrounding fat globules in milk. The purpose of this study was thus to identify MFGM proteins from Camelus dromedarius milk. Major MFGM proteins (namely, fatty acid synthase, xanthine oxidase, butyrophilin, lactadherin, and adipophilin) already evidenced in cow milk were identified in camel milk using mass spectrometry. In addition, a 1D-LC-MS/MS approach led us to identify 322 functional groups of proteins associated with the camel Milk Fat Globule Membrane. We hope that these findings will contribute to a better characterization of camel milk and to an improved understanding of lipid droplet formation in the mammary epithelial cell.
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Project description:The 24h gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes. Keywords: time course analysis
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Project description:The 4 day gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes. Serial milk samples were collected every 3h for 4 days from 5 normal, lactating women. RNA was isolated from the milk fat globules and utilized for microarray analyses.
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Project description:During lactation, mammary epithelial secretory cells secrete huge amounts of milk from their apical side. The major milk proteins, the caseins, are secreted by exocytosis, while milk fat globules are released by budding, enwrapped by the plasma membrane. Due to the number and large size of milk fat globules, the membrane surface needed for their secretion might exceed that of the apical plasma membrane. In order to identify the cellular compartments that may provide membrane during the budding of milk fat globules, a large-scale proteomics analysis of both cytoplasmic lipid droplets and secreted milk fat globules membranes was performed in mouse cells. The differential analysis of the protein profiles of these two organelles strongly suggest that, in addition to the apical plasma membrane, at least the endoplasmic reticulum, the secretory vesicles containing caseins, and potentially the mitochondria contribute to the formation of the milk fat globule membrane. Moreover, the specific analysis of the membrane-associated as well as the raft-associated proteins reinforces this possibility and points to a role for lipid rafts in milk product secretion. The subcellular localization of several major proteins of the cytoplasmic lipid droplets or of the milk fat globule membrane was investigated by immunofluorescence in lactating mammary epithelial cells. Furthermore, the localization of GM1 ganglioside, a known marker of lipid rafts and of free cholesterol, showed a clear association with both cytoplasmic lipid droplets and secreted milk fat globules. Additionally, the presence of some SNARE proteins that may be involved in casein exocytosis and of two Rab GTPases on both cytoplasmic lipid droplets and milk fat globules was demonstrated by immunofluorescence. Altogether, our results provide evidence for a pivotal role of the endoplasmic reticulum as membrane contributor to milk fat globule budding, and suggest that some SNARE proteins may spatio-temporally coordinate the secretion of milk products.
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Project description:Analysis of the milk fat globule proteome from mice with a mammary-specific knockout of Xanthine Oxidoreductase.
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