The Differential Composition of Whey Proteomes in Hu Sheep Colostrum and Milk during Different Lactation Periods
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Animals and Preparing of Whey Fractions
2.2. Two-Dimensional Gel Electrophoresis
2.3. Analysis of Gel Images and Protein Spots Abundance
2.4. In-Gel Digestion and Mass Spectrometric Analysis
2.5. Protein Identification
2.6. Bioinformatic Analysis
3. Results and Discussion
3.1. The Analysis of Two-Dimensional Electrophoresis Maps
3.2. Mass Spectrometry Analysis of Differentially Expressed Protein Spots
3.3. GO Annotation and KEGG Pathway Enrichment Analysis of DEPs
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Spot Number | D 0 | D 3 | D 7 | D 14 | D 28 | D 56 |
---|---|---|---|---|---|---|
1 | 256 | 156 | 0 | 100 | 2943 | 111 |
2 | 539 | 140 | 5297 | 135 | 2558 | 136 |
3 | 59,805 | 0 | 0 | 0 | 0 | 0 |
4 | 9070 | 0 | 0 | 0 | 0 | 0 |
5 | 9570 | 0 | 0 | 0 | 0 | 0 |
6 | 3908 | 156 | 0 | 0 | 0 | 0 |
7 | 12,886 | 107 | 0 | 125 | 644 | 0 |
8 | 6720 | 297 | 108 | 98 | 3495 | 102 |
9 | 15,387 | 393 | 3186 | 124 | 8728 | 137 |
10 | 10,252 | 732,945 | 6554 | 136 | 5530 | 0 |
11 | 9745 | 525 | 7490 | 153 | 8517 | 0 |
12 | 8298 | 136 | 8633 | 129 | 10,124 | 163 |
13 | 4143 | 0 | 2753 | 158 | 2659 | 0 |
14 | 10,893 | 139 | 0 | 0 | 0 | 0 |
15 | 13,823 | 223 | 0 | 0 | 0 | 0 |
16 | 12,456 | 144 | 0 | 0 | 0 | 0 |
17 | 8233 | 73 | 0 | 0 | 0 | 0 |
18 | 10,884 | 12 | 0 | 0 | 0 | 0 |
19 | 1450 | 222 | 0 | 110 | 1047 | 0 |
20 | 234 | 0 | 0 | 0 | 0 | 0 |
21 | 260 | 0 | 0 | 0 | 0 | 0 |
22 | 214 | 0 | 0 | 0 | 0 | 0 |
23 | 259 | 0 | 0 | 0 | 0 | 0 |
24 | 3597 | 45 | 1213 | 100 | 2992 | 0 |
25 | 1075 | 0 | 440 | 160 | 1233 | 0 |
26 | 1110 | 0 | 0 | 140 | 117 | 0 |
27 | 580 | 52 | 0 | 100 | 0 | 0 |
28 | 8724 | 0 | 0 | 0 | 1725 | 0 |
29 | 6494 | 0 | 0 | 0 | 2521 | 0 |
30 | 1152 | 0 | 633 | 112 | 0 | 0 |
31 | 390 | 0 | 0 | 0 | 0 | 0 |
32 | 3299 | 0 | 238 | 0 | 0 | 0 |
33 | 597 | 0 | 0 | 0 | 0 | 0 |
34 | 838 | 0 | 0 | 0 | 0 | 0 |
35 | 1834 | 0 | 0 | 0 | 0 | 0 |
36 | 2150 | 0 | 15,751 | 106 | 0 | 0 |
37 | 7599 | 321 | 0 | 100 | 0 | 110 |
38 | 6208 | 389 | 1046 | 100 | 11,750 | 100 |
39 | 3157 | 138 | 13,261 | 150 | 18,076 | 100 |
40 | 7160 | 139 | 3152 | 112 | 15,842 | 101 |
41 | 3134 | 11,587 | 13,087 | 11,570 | 2499 | 3568 |
42 | 0 | 5373 | 4586 | 5541 | 0 | 0 |
43 | 0 | 0 | 114 | 300 | 0 | 0 |
44 | 0 | 0 | 259 | 820 | 0 | 0 |
45 | 0 | 0 | 90 | 190 | 0 | 0 |
46 | 0 | 0 | 318 | 870 | 0 | 0 |
47 | 0 | 0 | 102 | 406 | 0 | 0 |
48 | 0 | 0 | 230 | 116 | 0 | 0 |
49 | 0 | 0 | 1125 | 532 | 0 | 0 |
50 | 0 | 0 | 283 | 305 | 0 | 0 |
51 | 386,070 | 1,207,960 | 638,265 | 581,040 | 529,120 | 500,480 |
52 | 1,190,595 | 457,725 | 283,400 | 274,480 | 156,200 | 25,240 |
Spot Number | Protein Name | Abbreviation | Organism | Accession No. | Protein MW | Protein PI | Pep. Count | Protein Score | Protein Score C. I. % | Total Ion | Total Ion C. I. % |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | Globin C, coelomic | GLBC | Ovis aries | tr|W5P5T4 | 103,838.6 | 5.87 | 17 | 163 | 100 | 121 | 100 |
2 | Protein yippee-like 5 (Fragment) | YPEL5 | Ovis aries | tr|W5QEE0 | 14,933.5 | 8.45 | 3 | 17 | 0 | ||
3 | Lactotransferrin | LTF | Capra hircus | sp|Q29477 | 79,361.3 | 6.75 | 21 | 205 | 100 | 158 | 100 |
4 | Lactotransferrin | LTF | Capra hircus | sp|Q29477 | 79,362.3 | 6.75 | 23 | 319 | 100 | 308 | 100 |
5 | Lactotransferrin | LTF | Capra hircus | sp|Q29477 | 79,363.3 | 6.75 | 21 | 123 | 100 | 78 | 100 |
6 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 13 | 154 | 100 | 122 | 100 |
7 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 14 | 375 | 100 | 337 | 100 |
8 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 14 | 392 | 100 | 354 | 100 |
9 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 12 | 387 | 100 | 359 | 100 |
10 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 16 | 450 | 100 | 401 | 100 |
11 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 13 | 377 | 100 | 345 | 100 |
12 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 13 | 378 | 100 | 346 | 100 |
13 | Polymeric immunoglobulin receptor | PIGR | Bos taurus | sp|P81265 | 83,694.6 | 7.07 | 15 | 206 | 100 | 162 | 100 |
14 | Immunoglobulin heavy constant mu | IGHM | Ovis aries | tr|W5NXW9 | 50,606.7 | 5.44 | 13 | 736 | 100 | 671 | 100 |
15 | Immunoglobulin heavy constant mu | IGHM | Ovis aries | tr|W5NXW9 | 50,606.7 | 5.44 | 12 | 521 | 100 | 468 | 100 |
16 | Immunoglobulin heavy constant mu | IGHM | Ovis aries | tr|W5NXW9 | 50,606.7 | 5.44 | 14 | 544 | 100 | 475 | 100 |
17 | Immunoglobulin heavy constant mu | IGHM | Ovis aries | tr|W5NXW9 | 50,606.7 | 5.44 | 11 | 467 | 100 | 423 | 100 |
18 | Immunoglobulin heavy constant mu | IGHM | Ovis aries | tr|W5NXW9 | 50,606.7 | 5.44 | 14 | 550 | 100 | 481 | 100 |
19 | Airway lactoperoxidase | ALPO | Ovis aries | tr|Q9MZY2 | 81,348.5 | 8.95 | 20 | 237 | 100 | 176 | 100 |
20 | Nucleobindin-1 | NUCB1 | Ovis aries | tr|W5PS94 | 53,546 | 5.13 | 18 | 137 | 100 | 68 | 99.9 |
21 | Nucleobindin-1 | NUCB1 | Ovis aries | tr|W5PS94 | 53,546 | 5.13 | 18 | 149 | 100 | 76 | 100 |
22 | Vitamin D-binding protein | VTDB | Bos taurus | sp|Q3MHN5 | 54,903.7 | 5.36 | 8 | 73 | 99.5 | 51 | 99.8 |
23 | Alpha-1-antitrypsin transcript variant 1 | A1ATV1 | Ovis aries | tr|I1WXR3 | 46,339.8 | 5.78 | 11 | 105 | 100 | 64 | 99.9 |
24 | Vitamin D-binding protein | VDBP | Bos taurus | sp|Q3MHN5 | 54,903.7 | 5.36 | 10 | 174 | 100 | 139 | 100 |
25 | Guanine nucleotide-binding protein subunit gamma | GNG5 | Ovis aries | tr|W5PWW2 | 7427.9 | 9.9 | 3 | 24 | 0 | ||
26 | Vimentin | VIM | Macaca fascicularis | sp|Q4R4X4 | 53,733.1 | 5.06 | 17 | 58 | 83.3 | ||
27 | Dual specificity phosphatase | DUPD | Gallus gallus | sp|P0C597 | 24,535.1 | 5.69 | 6 | 35 | 0 | ||
28 | Keratin 10 | KRT10 | Ovis aries | tr|W5Q160 | 57,476.7 | 5.4 | 12 | 197 | 100 | 158 | 100 |
29 | Keratin 10 | KRT10 | Ovis aries | tr|W5Q160 | 57,476.7 | 5.4 | 10 | 103 | 100 | 77 | 100 |
30 | Actin gamma 1 | ACTG1 | Ovis aries | tr|W5QAX3 | 42,292 | 5.31 | 23 | 775 | 100 | 613 | 100 |
31 | Actin gamma 1 | ACTG1 | Ovis aries | tr|W5QAX3 | 42,292 | 5.31 | 17 | 521 | 100 | 413 | 100 |
32 | Complement C3 | C3 | Bos taurus | sp|Q2UVX4 | 188,674.8 | 6.41 | 19 | 608 | 100 | 588 | 100 |
33 | Keratin 10 | KRT10 | Homo sapiens | sp|P13645 | 59,019.8 | 5.13 | 13 | 114 | 100 | 73 | 99.9 |
34 | G protein-regulated inducer of neurite outgrowth 1 | GPRININ1 | Ovis aries | tr|W5PH95 | 51,169.7 | 6.19 | 7 | 83 | 99.9 | 61 | 99.9 |
35 | G protein-regulated inducer of neurite outgrowth 1 | GPRININ1 | Ovis aries | tr|W5PH95 | 51,169.7 | 6.19 | 5 | 70 | 99.7 | 55 | 99.9 |
36 | Clusterin | CLU | Ovis aries | tr|W5PZI1 | 51,554.4 | 5.77 | 14 | 466 | 100 | 401 | 100 |
37 | Alpha s1 casein variant | CSN1S1 | Ovis aries | tr|D3TU01 | 23,469 | 5.64 | 10 | 511 | 100 | 454 | 100 |
38 | Alpha s1 casein variant | CSN1S1 | Ovis aries | tr|D3TU01 | 23,469 | 5.64 | 13 | 252 | 100 | 166 | 100 |
39 | Alpha s2 casein variant | CSN1S2 | Ovis aries | tr|D3TU01 | 23,469 | 5.13 | 15 | 323 | 100 | 271 | 100 |
40 | Kappa casein (Fragment) | CSN3 | Ovis aries | tr|A0A059T9V6 | 18,149 | 5.77 | 1 | 44 | 0 | 40 | 99.6 |
41 | Kappa-casein | CSN3 | Ovis aries | tr|A0A059T9N6 | 21,595.9 | 5.78 | 6 | 272 | 100 | 243 | 100 |
42 | Keratin 10 | KRT10 | Ovis aries | tr|W5Q160 | 57,476.7 | 5.4 | 11 | 251 | 100 | 220 | 100 |
43 | Glycosylation-dependent cell adhesion molecule 1 | GLYCAM1 | Ovis aries | tr|W5Q3I2| | 17,117 | 5.41 | 4 | 127 | 100 | 109 | 100 |
44 | Kinesin family member 20B | KIF20B | Ovis aries | tr|W5Q1Q1 | 211,015.5 | 5.45 | 39 | 66 | 99.2 | ||
45 | Kappa-casein | CSN3 | Ovis aries | tr|A0A059T9V6 | 21,595.9 | 5.78 | 4 | 75 | 99.8 | 55 | 99.9 |
46 | Kappa-casein | CSN3 | Ovis aries | tr|A0A059T9V6 | 21,595.9 | 5.78 | 4 | 156 | 100 | 141 | 100 |
47 | Kappa-casein | CSN3 | Ovis aries | tr|A0A059T9V6 | 21,595.9 | 5.78 | 4 | 92 | 99.9 | 73 | 100 |
48 | Kappa-casein | CSN3 | Ovis aries | tr|A0A059T9V6 | 21,595.9 | 5.78 | 5 | 152 | 100 | 131 | 100 |
49 | Kappa-casein | CSN3 | Ovis aries | tr|A0A059T9V6 | 21,595.9 | 5.78 | 4 | 135 | 100 | 117 | 100 |
50 | Uncharacterized protein | Ovis aries | tr|W5NPX0 | 22,587.1 | 5.29 | 4 | 20 | 0 | |||
51 | Serum albumin | ALB | Ovis aries | tr|W5PWE9 | 71,372.1 | 5.79 | 31 | 1020 | 100 | 822 | 100 |
52 | Immunoglobulin alpha heavy chain | IGHC | Ovis aries | tr|W5PH95 | 51,169.7 | 6.19 | 5 | 170 | 100 | 148 | 100 |
Pathway Name | Pathway ID | Count | p Value |
---|---|---|---|
Hippo signaling pathway * | oas04390 | 2 | 2.18 × 10−3 |
Arrhythmogenic right ventricular cardiomyopathy (ARVC) * | oas05412 | 1 | 3.46 × 10−2 |
Adherens junction * | oas04520 | 1 | 3.55 × 10−2 |
Viral myocarditis * | oas05416 | 1 | 3.89 × 10−2 |
Hypertrophic cardiomyopathy (HCM) * | oas05410 | 1 | 3.94 × 10−2 |
Bacterial invasion of epithelial cells * | oas05100 | 1 | 3.94 × 10−2 |
Dilated cardiomyopathy * | oas05414 | 1 | 4.23 × 10−2 |
Salmonella infection * | oas05132 | 1 | 4.38 × 10−2 |
Complement and coagulation cascades * | oas04610 | 1 | 4.76 × 10−2 |
Thyroid hormone signaling pathway | oas04919 | 1 | 5.58 × 10−2 |
Leukocyte transendothelial migration | oas04670 | 1 | 5.92 × 10−2 |
Platelet activation | oas04611 | 1 | 6.11 × 10−2 |
Tight junction | oas04530 | 1 | 6.73 × 10−2 |
Apoptosis | oas04210 | 1 | 7.20 × 10−2 |
Oxytocin signaling pathway | oas04921 | 1 | 7.72 × 10−2 |
Cell adhesion molecules (CAMs) | oas04514 | 1 | 7.91 × 10−2 |
Influenza A | oas05164 | 1 | 8.98 × 10−2 |
Phagosome | oas04145 | 1 | 9.12 × 10−2 |
Proteoglycans in cancer | oas05205 | 1 | 1.01 × 10−1 |
Rap1 signaling pathway | oas04015 | 1 | 1.02 × 10−1 |
Focal adhesion | oas04510 | 1 | 1.02 × 10−1 |
Regulation of actin cytoskeleton | oas04810 | 1 | 1.04 × 10−1 |
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Zhang, X.; Liu, X.; Li, F.; Yue, X. The Differential Composition of Whey Proteomes in Hu Sheep Colostrum and Milk during Different Lactation Periods. Animals 2020, 10, 1784. https://doi.org/10.3390/ani10101784
Zhang X, Liu X, Li F, Yue X. The Differential Composition of Whey Proteomes in Hu Sheep Colostrum and Milk during Different Lactation Periods. Animals. 2020; 10(10):1784. https://doi.org/10.3390/ani10101784
Chicago/Turabian StyleZhang, Xueying, Xinxin Liu, Fadi Li, and Xiangpeng Yue. 2020. "The Differential Composition of Whey Proteomes in Hu Sheep Colostrum and Milk during Different Lactation Periods" Animals 10, no. 10: 1784. https://doi.org/10.3390/ani10101784