Dataset Information

A proteomic characterization reveals differences in the milk fat globule membrane of buffalo and bovine milk

ABSTRACT: The proteins of the milk fat globule membrane (MFGM) have a number of functions, such as the regulation of milk fat secretion and metabolism, the uptake and transportation of fatty acids in the intestine and potential protection from bacterial or viral infection. While the proteome of the MFGM in bovine milk has been extensively characterised, our knowledge of these proteins in buffalo milk is limited. In this study, a proteomic approach was used to characterise the proteome of the buffalo MFGM. Multiple extraction techniques were employed to increase the coverage of proteins identified, while label free relative quantitative liquid chromatography tandem mass spectrometry was used for comparison between the buffalo and bovine MFGM proteome. A total of 220 buffalo MFGM proteins and 234 bovine MFGM proteins were identified after being filtered from the initial dataset of 757 and 680 proteins respectively. Significantly greater amounts of xanthine oxidoreductase, platelet glycoprotein 4, heat shock cognate and calcineurin B homologous protein were identified per mass of buffalo MFGM protein extracted. The higher expression of xanthine oxidase in the MFGM of buffalo milk was confirmed by Western blot analysis and a heterogeneous distribution of this protein observed in situ on the surface of the MFGM. The high concentration of fat in buffalo milk, together with the differences in the MFGM proteome indicate differences in the nutritional profile, biological function, and potential ease of processing of buffalo and bovine milk products.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Bos Taurus (bovine) Bubalus Bubalis

TISSUE(S): Milk Fat

SUBMITTER: Ching-Seng Ang

LAB HEAD: Sally L. Gras

PROVIDER: PXD005318 | Pride | 2017-05-19

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20130328_Hanh_BM_R1.msf	Msf
	20130328_Hanh_BM_R1.pep.pep.xml	Pepxml
	20130328_Hanh_BM_R1.raw	Raw
	20130328_Hanh_BM_R2.msf	Msf
	20130328_Hanh_BM_R2.pep.xml	Pepxml

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Publications

Proteomic characterization and comparison of mammalian milk fat globule proteomes by iTRAQ analysis.

Yang Yongxin Y Zheng Nan N Zhao Xiaowei X Zhang Yangdong Y Han Rongwei R Ma Lu L Zhao Shengguo S Li Songli S Guo Tongjun T Wang Jiaqi J

Journal of proteomics 20150108

Milk fat globule membrane (MFGM) proteins are known to be involved in many biological functions; however, their components and inter-species complexity have not yet been completely elucidated. We investigated the protein composition of the MFGM-enriched fraction from Holstein, Jersey, yak, buffalo, goat, camel, horse, and human. Extracted proteins from the MFGM-enriched fractions were identified and quantified by an iTRAQ proteomic approach. We identified 520 protein species categorized as biolo ...[more]

PMID: 25576853

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Project description:Studies of normal human mammary gland development and function have mostly relied on cell culture, limited surgical specimens, and rodent models. Although RNA extracted from human milk has been used to assay the mammary transcriptome non-invasively, the transcriptome derived from the milk fat layer has not been compared with the mammary-derived transcriptome nor have sources of RNA been quantified in milk. In this study the effects of milk collection and processing on RNA quality and origin were assessed in humans and rhesus macaques. Total RNA in milk was quantitated in acridine orange-stained milk using an automated whole slide scanner and custom-built Globulator software. Total RNA extracted from milk fat, cells in milk, and mammary biopsies of lactating rhesus macaques were compared using RNA sequencing and analysis. Compared with human milk, milk from macaques contained similar amounts of RNA-containing cytoplasmic crescents, but more cells. Total RNA extracted from milk fractions was also evaluated for factors that affect RNA quality. Degradation of RNA extracted from human milk fat was positively correlated with geographic distance from collection site, storage time, and sample type. There were no differences in RNA degradation in macaque milk collected after 10 min or 4 hr accumulation, suggesting that degradation of RNA extracted from milk fat may not occur in the mammary gland. Using RNA-Seq, RNA extracted from macaque milk fat and cells in milk more accurately represented RNA from mammary epithelial cells (cells that produce milk) than did RNA from mammary tissue. Mammary epithelium-specific transcripts were more abundant in macaque milk fat whereas adipose or stroma-specific transcripts were more abundant in mammary tissue. Functional analyses confirmed the validity of milk as a source of RNA from mammary epithelial cells. Analysis of highly abundant putative microRNAs in macaque milk fat revealed a potentially novel non-coding RNA species that is conserved in humans. RNA extracted from the milk fat during lactation accurately portrayed the RNA profile of milk-producing mammary epithelial cells. However, this sample type clearly requires protocols that minimize RNA degradation. Transcript profiles from milk cells, milk fat, and mammary tissue from 6 lactating rhesus macaques at 30 and 90 days lactation; 34 samples run in triplicate