Effect of Lactation on Proteome of Milk Fat Globule Membrane of Laoshan Dairy Goat
Description
We have investigated MFGM proteome of goat milk at 1, 3, 30, 90, 150, and 240 d using data-independent acquisition and data-dependent acquisition quantitative proteomics approaches. The clustering and principal component analysis revealed that the protein components in goat milk was associated with specific lactation stages. Protein-protein interaction showed that albumin had more interaction with other proteins and was considered as a central node. Our results can provide a better understanding of the MFGM proteome during the lactation cycle of dairy goats and indicate changed proteins are important for the better understanding of intrinsic physiological function of goats.
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DDA and DIA analyses were both performed on an EASY-nLC 1000 system coupled with Orbitrap Fusion Lumos (Thermo Fisher Scientific, Milford, MA, USA). The samples were loaded onto an Aqua C18 trapping column (100 μm × 2 cm, 5 μm; Thermo Fisher Scientific) with solution A (0.1% FA) at a flow rate of 300 nL/min and separated using an Aqua C18 analytical column (100 mm × 75 μm, 3 μm; Thermo Fisher Scientific). Gradient separation was performed by increasing the percentage of mobile phase B from 0% to 10% within 3 min, from 10% to 35% within 45 min, from 35% to 80% within 26 min, and to 100% in 1 min; final holding was performed at 100% for the final 15 min. For DDA analysis, mass spectrometry parameters were set as follows: MS scan range, 300−1800 m/z; normalized collision energy (NCE), 27 eV; MS resolution, 60,000; maximal injection time (MIT), 50 ms; RF lens, 30%; dynamic exclusion duration, 40 s; MS/MS high-energy collision dissociation (HCD) scan resolution, 15,000; MIT, 50 ms; and automated gain control (AGC) target, 50,000. For DIA analysis, the same nano-LC system and gradient as in the DDA analysis were used. The DIA MS parameters were set as follows: MS scan range, 395−1205 m/z; NCE, 30; MS resolution, 60,000; MIT, 100 ms; MS/MS HCD scan resolution, 15,000; and AGC target, 1e6.