Abstract
A fertilized chicken egg is a unit of life. During hatching, transport of nutrients, including calcium, have been reported from the egg components to the developing embryo. Calcium is mobilized from the eggshell with the involvement of Ca2+-binding proteins. In addition, other unknown proteins may also play some important roles during embryo developing process. Therefore identification and prediction of biological functions of eggshell membrane (ESM) proteins during chick embryo development was conducted by proteome analysis. Comparison of different lysis solutions indicated that the highest ability to extract ESM proteins could be obtained with 1 % sodium dodecyl sulfate in 5 mM Tris–HCl buffer pH 8.8 containing 0.1 % 2-mercaptoethanol. In this study fertilized Cornish chicken eggs were incubated at 37 °C in humidified incubators for up to 21 days. At selected times (days 1, 9, 15 and 21), samples were taken and the ESMs were carefully separated by hand, washed with distilled water, and air-dried at room temperature. The ESM proteins were then solubilized and analyzed by proteome analysis. Sodium dodecyl sulfate polyacrylamide gel electrophoresis combined with high performance liquid chromatography and mass spectrometry revealed 62 proteins in the ESM; only keratin is known ESM protein, 8 of which are egg white proteins and related while 53 others have not previously been reported. Some differences in the types of proteins and their molecular functions were noted in ESM at different incubation times. One protein which was present only at days 15 and 21 of egg incubation was identified as a calcium binding protein i.e. EGF like repeats and discoidin I like domain 3 (EDIL3 homologous protein).
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Abbreviations
- ACN:
-
Acetonitrile
- BSA:
-
Bovine serum albumin
- ESM:
-
Eggshell membrane
- ESI/Q-TOF:
-
Electrospray ionization/quadupole-time of fight
- GOCat:
-
Gene ontology categorizer
- LC-MS–MS:
-
High performance liquid chromatography–Tandem mass spectrometry
- β-NAGase:
-
β-N-acetylglucosaminidase
- NCBI:
-
National center for biotechnology information
- SDS:
-
Sodium dodecyl sulfate
- SDS–PAGE:
-
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
References
Ahlborn GJ, Clare DA, Sheldon BW, Kelly RW (2006) Prot J 25:71–81
Akins RE, Tuan RS (1993) J Cell Sci 105:369–379
Arias JL, Fernandez MS, Dennis JE, Caplan AI (1991) Connect Tissue Res 26:37–41
Baker JR, Balch DA (1962) Biochem J 82:352–361
Balcerzak M, Malinowska A, Thouverey C, Sekrecka A, Dadlez M, Buchet R, Pikula S (2008) Proteomics 8:192–205
Bingle CD, Craven J (2004) Trends Immunol 25(2):53–55
Brockway BE, Forster SJ, Freedman RB (1980) Biochem J 191:873–876
Crooks RJ, Simkiss K (1974) J Exp Biol 61:197–202
Daengprok W, Garnjanagoonchorn W, Naivikul O, Pornsinlpatip P, Issigonis K, Mine Y (2003) J Agric Food Chem 51:6056–6061
Denzer AJ, Gesemann M, Schumacher B, Ruegg MA (1995) J Cell Biol 31:1547–1560
Ellis JH, Richards DE, Rogers JH (1991) Cell Tissue Res 264:197–208
Elsbach P, Weiss J (1998) Curr Opin Immunol 10(45):45–49
Fernandez MS, Araya M, Arias JL (1997) Matrix Biol 16:13–20
Gettins P (2002) Chem Rev 102(12):4751–4804
Gillepie JI, Green Well JR (1988) J Physiol 405:385–395
Gromov P, Gromova I, Celis JE (2006) In: Veenstra TD, Yates JR (eds) Proteomics for biological discovery. Wiley, New Jersey
Guearin-Dubiard C, Pasco M, Mollea D, Deasert C, Croguennec T, Nau F (2006) J Agri Food Chem 54:3901–3910
Hamburger V, Hamilton HL (1951) J Morphol 88:49–92
Heckler EJ, Rancy PC, Kodali VK, Thorpe C (2008) Biochi Bioph Acta 1783:567–577
Hernandez-Hernandez A, Vidal ML, Gomez-Morales J, Rodriguez-Navarro AB, Labas V, Gautron J, Nys Y, Garcia Ruiz JM (2008) J Cryst Grow 310:1754–1759
Hinckea MT, Gautron UJ, Panheleux M, Garcia-Ruizc J, McKeed MD, Nysb Y (2000) Matrix Biol 19:443–453
Johnston PM, Comar CL (1955) Am J Physiol 183:365–370
Jost JP (1993) J Proc Natl Acad Sci USA 89:4684–4688
Kodali VK, Gannon SA, Paramasivam S, Raje S, Polenova T, Thorpe C (2000) PLoS One 6(3):e18187
Laemmli UK (1970) Nature 227:680–685
Liu S, Piwnica-Worms D, Lieberman M (1990) J Gene Physilo 96:1247–1269
Liu S, Wang L, Wang N, Wang Y, Shi H, Li H (2009) Biochem Physiol A Mol Integr Physiol 154:135–141
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) J Biol Chem 193:265–275
Moreira EF, Adler R (2006) Dev Biol 298:272–284
Nakano T, Ikawa NI, Ozimek L (2003) Poult Sci 82:510–514
Nys Y, Gautron J, Garcia-Ruiz JM, Hincke MT (2004) C. R. Palevol 3:549–562
Rabilloud T (2006) In: Veenstra TD, Yates JR (eds) Proteomics for biological discovery. Wiley, New Jersey
Scances CG, Brant G, Ensminger ME (2004) Poultry science. Pearson Education, Inc., New Jersey
Simkiss K (1961) Biol Rev Camb Philos Soc 36:312–367
Simkiss K (1967) Nature 214:84–86
Takahashi K, Shirai K, Kitamura M, Hattori M (1996) Biosci Biotechnol Biochem 60:1299–1302
Tuan RS, Carson MJ, Jozefiak JA (1986) J Cell Sci 82:73–84
Vanhoutteghem A, Londero T, Ghinea N, Djian P (2004) Differentiation 72:123–137
Whisstock J, Skinner R, Lesk AM (1998) Trends Biochem Sci 23(2):63–67
Wong M, Hendrix MJC, von der Mark K, Little C, Stern R (1984) Dev Biol 104:28–36
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The authors thank The BIOTEC Genome Institute (National Science and Technology Development Agency, Thailand) for their facilities. The support fund for the study was provided by the Graduate School, Kasetsart University.
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Kaweewong, K., Garnjanagoonchorn, W., Jirapakkul, W. et al. Solubilization and Identification of Hen Eggshell Membrane Proteins During Different Times of Chicken Embryo Development Using the Proteomic Approach. Protein J 32, 297–308 (2013). https://doi.org/10.1007/s10930-013-9487-0
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DOI: https://doi.org/10.1007/s10930-013-9487-0