Dataset Information

Milk glycopeptide across lactation

ABSTRACT: Protein N-glycosylation is an essential post-translational modification (PTM) frequently occurring on human milk proteins. These PTMs assist in protecting the infant's health and function amongst others as competitive inhibitors of pathogen binding and immunomodulators. Due to the individual uniqueness of each mother’s milk and the overall complexity and temporal changes of protein N-glycosylation, analysis of the human milk N-glycoproteome requires longitudinal personalized approaches, providing protein- and N-site-specific quantitative information. Here we describe an automated platform using HILIC-based cartridges enabling the proteome-wide monitoring of N-glycopeptides using just 150 μg of breast milk protein digestion. Reproducibility in enrichment and quantitation is demonstrated in triplicate measurements of milk samples taken along an extended lactation period. Using higher energy c-trap dissociation (HCD) triggered electron-transfer higher-energy collision dissociation (EThcD), we were able to map around 1700 glycopeptides from 110 glycoproteins occurring on 191 glycosites, of which 43 sites were not before reported with experimental evidence. We next quantified many of these glycopeptides using a targeted proteomics approach with scheduled selective ion monitoring (SIM) and parallel reaction monitoring (PRM). In a 90 min gradient, we could quantitatively monitor 318 glycopeptides originating from 51 glycoproteins. Although each glycoprotein, each N-glycosylation site and each glycan attached revealed its own dynamic changes, we did observe a few general trends. Overall fucosylation especially terminal fucosylation increased across the lactation period.

ORGANISM(S): Homo Sapiens

SUBMITTER: Jing Zhu

PROVIDER: PXD022415 | panorama | Sat Nov 07 00:00:00 GMT 2020

REPOSITORIES: PanoramaPublic

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Publications

Quantitative Longitudinal Inventory of the <i>N</i>-Glycoproteome of Human Milk from a Single Donor Reveals the Highly Variable Repertoire and Dynamic Site-Specific Changes.

Zhu Jing J Lin Yu-Hsien YH Dingess Kelly A KA Mank Marko M Stahl Bernd B Heck Albert J R AJR

Journal of proteome research 20200327 5

Protein N-glycosylation on human milk proteins assists in protecting an infant's health and functions among others as competitive inhibitors of pathogen binding and immunomodulators. Due to the individual uniqueness of each mother's milk and the overall complexity and temporal changes of protein N-glycosylation, analysis of the human milk <i>N</i>-glycoproteome requires longitudinal personalized approaches, providing protein- and N-site-specific quantitative information. Here, we describe an aut ...[more]

PMID: 32125861

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Project description:A better understanding of impacts of nutrition during pre- weaning and post- weaning period is needed in order to design feeding strategies that optimize lifetime productivity. Therefore, the objectives of the present study were three-fold: to evaluate nutrition-induced changes of 1) growth from birth to first calving in heifers, 2) its impact on first-lactation milk yield and composition and 3) evaluate nutrition-induced changes in gene expression of mammary gland in Parda de Montaña heifers. Sixteen Parda de Montaña heifers, were randomly but equally assigned at birth to one of four management strategies, in a 2 x 2 factorial experiment. In the pre-weaning period (0-6 months of age), all the heifers suckled their dams twice daily for 30 minutes. Half of the heifers (n=8) were supplemented a starter concentrate (CON) while the other half only had access to their dam’s milk (NOCON). In the post-weaning period (6-15 months), half of the heifers of each pre-weaning feeding treatment received alfalfa hay ad libitum plus concentrates, with a different forage to concentrate ratio (F:C), 67:33 (67F:33C) or 87:13 (87F:13C). Plasma metabolites and hormones concentration at weaning was determined, and cows and their calves weigh was recorded once/week throughout the first lactation, and their ADG was calculated. Milk composition and quantity was determined during the first 4 months of lactation. Mammary tissue samples were taken by biopsy at weaning at the end of first lactation. RNA mammary gland samples (n=16, 8 samples from each treatment) were analyzed by using Bovine Gene 1.1 ST Array Strip. The results of the present study showed that heifers that received concentrate during pre- weaning period had greater BW at weaning when compared to NOCON heifers. Concentrate supplementation during the pre-weaning period increased heifers’ weight gains in the pre-weaning period (P < 0.001) but decreased weight gains in the post-weaning period (P < 0.01) and during pregnancy (P < 0.05). In addition, CON cows had lower milk production throughout the 4 months of the first lactation when compared with NOCON (P < 0.01). Moreover CON cows had decreased milk crude protein, crude fat, lactose, extract and casein content in their first lactation. In addition, cows that received concentrate during pre- weaning period showed greater SCC at the third and fourth month of lactation. Different gene expression patterns were found between CON and NOCON animals. Overall, concentrate supplementation at pre- weaning resulted in up –regulation of genes related with immune response and chemokine activity, suggesting that these animals might be in a compromised immune status, probably being at greater risk to develop mastitis in the later periods.

Dataset Information

Milk glycopeptide across lactation

Publications

Quantitative Longitudinal Inventory of the <i>N</i>-Glycoproteome of Human Milk from a Single Donor Reveals the Highly Variable Repertoire and Dynamic Site-Specific Changes.

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