Quantitative label-free site-specific glycoproteomic analysis of the milk fat globule membrane protein in human colostrum and mature milk

Guan, Boyuan; Zhang, Zhenghan; Liu, Xiaoyu; Song, Zhao; Bai, Xue; Luo, Xue; Feng, Daguang; Liu, Yang; Cao, Xiaohong; Yue, Xiqing

doi:10.1016/j.carbpol.2023.120588

Cited by 9 publications

(12 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MFGM protein extraction was performed according to a previously published protocol with minor adjustments. 13 Donkey milk (15 mL) was thawed at 4 °C and centrifuged at 8000g for 30 min to separate the upper milk fat layer. The milk fat fraction was added to a 6-fold volume of prechilled phosphate buffered saline solution (pH 7.4) and centrifuged at 8000g and 4 °C for 25 min; this washing procedure was repeated five times.…”

Section: Milk Sample Collectionmentioning

confidence: 99%

Comparative Site-Specific O-Glycosylation Analysis of the Milk Fat Globule Membrane Proteome in Donkey Colostrum and Mature Milk

Guan,

Cao,

Yang

et al. 2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

Donkey milk fat globule membrane (MFGM) proteins are a class of membrane-bound secreted proteins with broadspectrum biofunctional activities; however, their site-specific O-glycosylation landscapes have not been systematically mapped. In this study, an in-depth MFGM O-glycoproteome profile of donkey milk during lactation was constructed based on an intact glycopeptide-centered, label-free glycoproteomics pipeline, with 2137 site-specific O-glycans from 1121 MFGM glycoproteins and 619 site-specific O-glycans from 217 MFGM glycoproteins identified in donkey colostrum and donkey mature milk, respectively. As lactation progressed, the number of site-specific O-glycans from three glycoproteins significantly increased, whereas that of 11 sitespecific O-glycans from five glycoproteins significantly decreased. Furthermore, donkey MFGM O-glycoproteins with core-1 and core-2 core structures and Lewis and sialylated branch structures may be involved in regulating apoptosis. The findings of this study reveal the differences in the composition of donkey MFGM O-glycoproteins and their site-specific O-glycosylation modification dynamic change rules during lactation, providing a molecular basis for understanding the complexity and biological functions of donkey MFGM protein O-glycosylation.

show abstract

Section: Milk Sample Collectionmentioning

confidence: 99%

Comparative Site-Specific O-Glycosylation Analysis of the Milk Fat Globule Membrane Proteome in Donkey Colostrum and Mature Milk

Guan,

Cao,

Yang

et al. 2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The milk protein also contains a group of proteins associated with the milk fat globule membrane, which are distributed in the MFGM in an asymmetric form and account for approximately 1–4% of the total milk protein. , In addition, MFGM is glycosylated and phosphorylated, owing to its role as a membrane protein. To date, proteomics, glycoproteomics, and phosphorylation proteomics of MFGM proteins have revealed the abundance of MFGM proteins and glycoproteins and phosphorylated MFGM proteins in human, donkey, and bovine milk. ,,,− In addition, studies have shown that the concentration and quantity of MFGM vary continuously during lactation, with higher levels of MFGM in colostrum than in mature milk.…”

Section: Proteome-based Investigation Of the Effects Of Protein Analy...mentioning

confidence: 99%

“…There have also been some studies on the changes in human whey and MFGM proteins following post-translational modifications during lactation. Label-free glycoproteomics was used to identify 2617 and 986 unique site-specific N-glycans from 221 and 200 MFGM glycoproteins in DC and donkey mature milk (DM), respectively . Phosphorylation modifications of MFGM proteins in human milk have also been investigated.…”

Section: Proteome-based Investigation Of the Effects Of Protein Analy...mentioning

confidence: 99%

“…Label-free glycoproteomics was used to identify 2617 and 986 unique site-specific N-glycans from 221 and 200 MFGM glycoproteins in DC and donkey mature milk (DM), respectively. 76 Phosphorylation modifications of MFGM proteins in human milk have also been investigated. In human colostrum and mature milk, 305 phosphorylation sites from 170 phosphoproteins and 269 phosphorylation sites from 170 phosphoproteins have been quantified using untargeted phosphoproteomics.…”

Section: Advances In Human Milkmentioning

confidence: 99%

“…Previous studies have identified phosphorylation sites in α S1 -CN on S 56 , S 61 , S 63 , S 79 , S 81 , S 82 , S 83 , S 90 , and S 130 . In α S2 -CN, the identified phosphorylation sites are S 23 , S 24 , S 25 , S 31 , S 46 , S 71 , S 72 , S 73 , S 76 , and T 81 . β-CN has been found to be phosphorylated at S 144 , T 145 , S 146 , and S 158 .…”

Section: Advances In Cow Milk Protein Studiesmentioning

confidence: 99%

See 2 more Smart Citations

Proteomics and Peptidomics As a Tool to Compare the Proteins and Endogenous Peptides in Human, Cow, and Donkey Milk

Ning,

Yang,

Liu

et al. 2023

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

Cow’s milk is the most widely used ingredient in infant formulas. However, its specific protein composition can cause allergic reactions. Finding alternatives to replace cow’s milk and fill the nutritional gap with human milk is essential for the health of infants. Proteomic and peptidomic techniques have supported the elucidation of milk’s nutritional ingredients. Recently, omics approaches have attracted increasing interest in the investigation of milk because of their high throughput, precision, sensitivity, and reproducibility. This review offers a significant overview of recent developments in proteomics and peptidomics used to study the differences in human, cow, and donkey milk. All three types of milks were identified to have critical biological functions in human health, particularly in infants. Donkey milk proteins were closer in composition to human milk, were less likely to cause allergic reactions, and may be developed as novel raw materials for formula milk powders.

show abstract

Analysis of N‐ and O‐linked site‐specific glycosylation by ion mobility mass spectrometry: State of the art and future directions

Girgis,

Petruncio,

Russo

et al. 2024

Proteomics

View full text Add to dashboard Cite

Glycosylation, the major post‐translational modification of proteins, significantly increases the diversity of proteoforms. Glycans are involved in a variety of pivotal structural and functional roles of proteins, and changes in glycosylation are profoundly connected to the progression of numerous diseases. Mass spectrometry (MS) has emerged as the gold standard for glycan and glycopeptide analysis because of its high sensitivity and the wealth of fragmentation information that can be obtained. Various separation techniques have been employed to resolve glycan and glycopeptide isomers at the front end of the MS. However, differentiating structures of isobaric and isomeric glycopeptides constitutes a challenge in MS‐based characterization. Many reports described the use of various ion mobility–mass spectrometry (IM–MS) techniques for glycomic analyses. Nevertheless, very few studies have focused on N‐ and O‐linked site‐specific glycopeptidomic analysis. Unlike glycomics, glycoproteomics presents a multitude of inherent challenges in microheterogeneity, which are further exacerbated by the lack of dedicated bioinformatics tools. In this review, we cover recent advances made towards the growing field of site‐specific glycosylation analysis using IM–MS with a specific emphasis on the MS techniques and capabilities in resolving isomeric peptidoglycan structures. Furthermore, we discuss commonly used software that supports IM–MS data analysis of glycopeptides.

show abstract

Quantitative label-free site-specific glycoproteomic analysis of the milk fat globule membrane protein in human colostrum and mature milk

Cited by 9 publications

References 37 publications

Comparative Site-Specific O-Glycosylation Analysis of the Milk Fat Globule Membrane Proteome in Donkey Colostrum and Mature Milk

Comparative Site-Specific O-Glycosylation Analysis of the Milk Fat Globule Membrane Proteome in Donkey Colostrum and Mature Milk

Proteomics and Peptidomics As a Tool to Compare the Proteins and Endogenous Peptides in Human, Cow, and Donkey Milk

Analysis of N‐ and O‐linked site‐specific glycosylation by ion mobility mass spectrometry: State of the art and future directions

Contact Info

Product

Resources

About