Interrogation of Milk-Driven Changes to the Proteome of Intestinal Epithelial Cells by Integrated Proteomics and Glycomics

Abstract

Bovine colostrum is a rich source of bioactive components which are important in the development of the intestine, in stimulating gut structure and function and in preparing the gut surface for subsequent colonization of microbes. What is not clear, however, is how colostrum may affect the repertoire of receptors and membrane proteins of the intestinal surface and the post-translational modifications associated with them. In the present work, we aimed to characterize the surface receptor and glycan profile of human HT-29 intestinal cells after exposure to a bovine colostrum fraction (BCF) by means of proteomic and glycomic analyses. Integration of label-free quantitative proteomic analysis and lectin array profiles confirmed that BCF exposure results in changes in the levels of glycoproteins present at the cell surface and also changes to their glycosylation pattern. This study contributes to our understanding of how milk components may regulate intestinal cells and prime them for bacterial interaction.

Comprehensive transcriptomic and proteomic analyses identify intracellular targets for myriocin to induce Fusarium oxysporum f. sp. niveum cell death

“…Next, lysis buffer (8 M urea, 1% SDS, and 1% protease inhibitor) was used to redissolve the protein precipitate, and the lysates were centrifuged; the protein concentration in the supernatant was quantified by using the bicinchoninic acid (BCA) method. Protein digestion was conducted following a standard procedure previously described by Morrin et al [60]. Briefly, the appropriate amount of trichloroethyl phosphate was mixed with 150 μg of protein in each sample tube, and the mixtures were incubated at 37°C for 60 min.…”

“…The sample was resuspended in 150 μL of buffer (100 mM TEAB). Trypsin was mixed with approximately 150 μg of protein for each sample (enzyme:protein, 1:50) and digested at 37°C overnight [60]. After trypsin digestion, the peptides were desalted using OASIS ® HLB μElution plates and vacuum dried, and the peptide concentrations were estimated using a peptide quantification kit (Thermo, Cat.…”

Bifidobacterium longum subsp. infantis ATCC 15697 and Goat Milk Oligosaccharides Show Synergism In Vitro as Anti-Infectives against Campylobacter jejuni

“…The HT-29 cells are extensively used as a model of the gastrointestinal tract, particularly as in vitro intestinal models of bacterial colonisation [41,42]. These cells exhibit classical characteristics that model small intestinal absorptive epithelial cells upon reaching confluence [43] and are a useful indicator of the structural landscape of the intestinal epithelium [44]. In the current study, we also employed HT-29 cells and hypothesised that the increase in B. infantis adhesion following GMO treatment may provide a protective effect against C. jejuni colonisation of HT-29 cells.…”

Human breast milk enhances intestinal mucosal barrier function and innate immunity in a pediatric human enteroid model

“…Bovine colostrum has been shown to influence the proteome of HT-29 cells as well as epithelial cell glycosylation (72). We show, for the first time, that human milk influences the synthesis of multiple mediators of metabolic and physiologic functions that act locally or systemically.…”