Proteomics-Driven Analysis of Ovine Whey Colostrum

Scumaci, Domenica; Trimboli, Francesca; Dell’Aquila, Ludovica; Concolino, Antonio; Pappaianni, Giusi; Tammè, Laura; Vignola, G.; Luciani, A; Morelli, Daniela; Cuda, Giovanni; Boari, Andrea; Britti, Domenico

doi:10.1371/journal.pone.0117433

Cited by 23 publications

(21 citation statements)

References 56 publications

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“…The above factors reduced the number of proteins but resulted in a protein detection level comparable to that of previous studies (Zhang et al, 2015). Some exceptions to this postulate are the finding of 342 proteins found in the fluid phase of ovine colostrum (Scumaci et al, 2015) and of 1,606 proteins Table 1. The proteins in the top right groups, the milk fluid phase proteins only (12 and 1), are presented in Table 2.…”

Section: Total Detection Of Proteins By 2d-lc-ms/mssupporting

confidence: 54%

Colostrum and milk protein rankings and ratios of importance to neonatal calf health using a proteomics approach

Nissen

Andersen

Bendixen

et al. 2017

Journal of Dairy Science

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Administration of colostrum to the newborn calf before gut closure is pivotal to its health, because of the transfer of passive immunity. Traditionally, passive immunity has been attributed to the transfer of immunoglobulins although it is increasingly clear that multiple other factors contribute, including innate immune proteins, developmental factors, immunomodulatory factors, and the presence of cellular immunity. The objective of this study was to produce a comprehensive comparison of the bovine colostrum proteome and the milk proteome by applying 2-dimensional liquid chromatography-tandem mass spectrometry. Further, the objectives were to rank proteins mutually and generate protein ratios from the spectral counts of the 2 proteomes and ELISA to gain insight into which proteins could be of most relevance to neonatal calf health. To obtain an in-depth picture of the bovine colostrum and milk proteome, we compared the contents of different fractions from bovine colostrum and milk from our 2 previous studies. A total of 140 colostrum fluid-phase proteins and 103 milk fluid-phase proteins were detected. In the cellular fraction, 324 and 310 proteins were detected in colostrum and milk, respectively. In total, 514 proteins were detected, of which 162 were in the fluid phase. Of these, 50 proteins were exclusively seen in colostrum, 13 were exclusively seen in milk, and 99 were common to colostrum and milk. Ranking of proteins mutually and calculating protein ratios based on spectral counts and ELISA resulted in new information on how proteins were associated with the fluid or cellular fraction of the samples. Interestingly, despite lower counts/concentrations than the classical proteins such as immunoglobulins, β-lactoglobulin, and lactotransferrin, several proteins appeared in higher or similar colostrum:milk spectral count ratios as these. Using this approach indicated, for example, that osteopontin, haptoglobin, milk amyloid A, and gelsolin may be interesting molecules to study in detail in their relation to calf health. Although the sensitivity, identification, and ranking of proteins varied between the 2 methods, and proteome analysis clearly suffers from low sensitivity, we believe that this idea and approach of generating ratios and ranking proteins can contribute new information and perspectives on how to prioritize the importance of multiple proteins, beyond immunoglobulins, in relation to neonatal calf health.

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Section: Total Detection Of Proteins By 2d-lc-ms/mssupporting

confidence: 54%

Colostrum and milk protein rankings and ratios of importance to neonatal calf health using a proteomics approach

Nissen

Andersen

Bendixen

et al. 2017

Journal of Dairy Science

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“…The large dynamic range of protein abundance has hindered conventional protein identification approaches, such as ion exchange chromatography and 2D-PAGE coupled with mass spectrometry. A recent proteomic study of whey from sheep colostrum (milk produced during late pregnancy) using 1D-PAGE coupled with LCMS/MS identified 343 proteins [ 31 ]. In a previous proteomic study comparing human colostrum with late lactation human milk considerable differences in protein complement were found [ 18 ], indicating the important role played by colostrum in early infant development.…”

Section: Resultsmentioning

confidence: 99%

In-Depth Characterization of Sheep (Ovis aries) Milk Whey Proteome and Comparison with Cow (Bos taurus)

Sabherwal

Duncan

et al. 2015

PLoS ONE

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An in-depth proteomic study of sheep milk whey is reported and compared to the data available in the literature for the cow whey proteome. A combinatorial peptide ligand library kit (ProteoMiner) was used to normalize protein abundance in the sheep whey proteome followed by an in-gel digest of a 1D-PAGE display and an in-solution digestion followed by OFFGEL isoelectric focusing fractionation. The peptide fractions obtained were then analyzed by LC-MS/MS. This enabled identification of 669 proteins in sheep whey that, to our knowledge, is the largest inventory of sheep whey proteins identified to date. A comprehensive list of cow whey proteins currently available in the literature (783 proteins from unique genes) was assembled and compared to the sheep whey proteome data obtained in this study (606 proteins from unique genes). This comparison revealed that while the 233 proteins shared by the two species were significantly enriched for immune and inflammatory responses in gene ontology analysis, proteins only found in sheep whey in this study were identified that take part in both cellular development and immune responses, whereas proteins only found in cow whey in this study were identified to be associated with metabolism and cellular growth.

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“…In addition to a-lact, defense-related proteins, and IGF-II/IGF-binding proteins, more than 100 proteins were identified in the milk of the Yangtze finless porpoise (Table S1). Many of the proteins in this study were also recently reported in the milk of animals and humans (D'Alessandro et al 2010, Ogawa et al 2014, Ha et al 2015, Scumaci et al 2015, Yang et al 2015. In addition to being a source of amino acids for newborns (L€ onnerdal 2003), the physiological functions of most milk proteins remain uncertain (Table S2) and thus require in vivo experiments to verify.…”

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confidence: 71%

“…The milk samples were immediately frozen in a liquid-nitrogen dewar and then stored at 280 C in a laboratory freezer until analysis. Milk whey samples were prepared according to the protocol described by Scumaci et al (2015), with modification of the low-speed centrifugation to 10 min. The total protein concentrations of the whey samples were determined by a BCA protein assay kit P0012 (Beyotime Institute of Biotechnology, Shanghai, China) prior to protein digestion.…”

mentioning

confidence: 99%