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The Protein Journal

, Volume 37, Issue 1, pp 21–37 | Cite as

Analysis of Siamese Crocodile (Crocodylus siamensis) Eggshell Proteome

  • Ivan Mikšík
  • Statis Paradis
  • Adam Eckhardt
  • David Sedmera
Article

Abstract

The proteins and pigment of the eggshell of the Siamese crocodile (Crocodylus siamensis) were analysed. For proteomic analysis, various decalcification methods were used when the two main surface layers were analyzed. These layers are important for antimicrobial defense of egg (particularly the cuticle). We found 58 proteins in both layers, of which 4 were specific for the cuticle and 26 for the palisade (honeycomb) layer. Substantial differences between proteins in the eggshell of crocodile and previously described birds’ eggshells exist (both in terms of quality and quantity), however, the entire proteome of Crocodilians has not been described yet. The most abundant protein was thyroglobulin. The role of determined proteins in the eggshell of the Siamese crocodile is discussed. For the first time, the presence of porphyrin pigment is reported in a crocodilian eggshell, albeit in a small amount (about 2 to 3 orders of magnitude lower than white avian eggs).

Keywords

Proteins of eggshell Eggshell pigment Crocodile 

Notes

Acknowledgements

We greatly appreciate the donation of crocodile’s eggshells by Ing. Procházka from the Crocodile ZOO Protivín. Financial support from the Czech Science Foundation (GA CR) (15-01948S - IM, SP, AE and 16-02972S - DS) is gratefully acknowledged.

Compliance with Ethical Standards

Conflict of interest

All authors declare that there are no competing interests.

Supplementary material

10930_2017_9750_MOESM1_ESM.pdf (124 kb)
Supplementary material 1 (PDF 124 KB)

References

  1. 1.
    Arias JL, Wiff JP, Fuenzalida V, Fernandez MS (2003) Molecular Regulation of Avian Eggshell Biomineralization. In: Kobayashi I, Ozawa H (eds) Biomineralization: formation, diversity, evolution and application. Tokai University Press, Japan, pp 13–17Google Scholar
  2. 2.
    Miksik I, Sedlakova P, Lacinova K, Pataridis S, Eckhardt A (2010) Determination of insoluble avian eggshell matrix proteins. Anal Bioanal Chem 397:205–214CrossRefGoogle Scholar
  3. 3.
    Ferguson M.W.J. (1985) Reproductive biology and embryology of the crocodilians. In: Gans C (ed) Biology of reptilia. vol 14A, Wiley, New York, pp 329–491Google Scholar
  4. 4.
    Packard MJ, DeMarco VG, Packard MJ, DeMarco. VG (1991) Eggshell structure and formation in eggs of oviparous reptiles. In: Deeming DC, Ferguson M.W.J. (eds) Egg incubation: its effects on embryonic development in birds and reptiles. Cambridge University Press, Cambridge, pp 53–69CrossRefGoogle Scholar
  5. 5.
    Prajanban B-O, Shawsuan L, Daduang S, Kommanee J, Roytrakul S, Dhiravisit A, Thammasirirak S (2012) Identification of five reptile egg whites protein using MALDI-TOF mass spectrometry and LC/MS-MS analysis. J Proteom 75:1940–1959CrossRefGoogle Scholar
  6. 6.
    Chaipayang S, Heamatorn N, Keha L, Daduang S, Songsiriritthigul C, Swatsitang P, Dhiravisit A, Thammasirirak S (2013) Purification and characterization of ovotransferrin from Crocodylus siamensis. Protein J 32:89–96CrossRefGoogle Scholar
  7. 7.
    Mann K, Macek B, Olsen JV (2006) Proteomic analysis of the acid-soluble organic matrix of the chicken calcified eggshell layer. Proteomics 6:3801–3810CrossRefGoogle Scholar
  8. 8.
    Mann K, Olsen JV, Maček B, Gnad F, Mann M (2008) Identification of new chicken egg proteins by mass spectrometry-based proteomic analysis. World’s Poult Sci J 64:209–218CrossRefGoogle Scholar
  9. 9.
    Farinazzo A, Restuccia U, Bachi A, Guerrier L, Fortis F, Boschetti E, Fasoli E, Citterio A, Righetti PG (2009) Chicken egg yolk cytoplasmic proteome, mined via combinatorial peptide ligand libraries. J Chromatogr A 1216:1241–1252CrossRefGoogle Scholar
  10. 10.
    Ahmed TAE, Suso H-P, Hincke MT (2017) In-depth comparative analysis of the chicken eggshell membrane proteome. J Proteom 155:49–62CrossRefGoogle Scholar
  11. 11.
    Mann K, Mann M (2013) The proteome of the calcified layer organic matrix of Turkey (Meleagris gallopavo) eggshell. Proteome Sci 11:40CrossRefGoogle Scholar
  12. 12.
    Mann K, Mann M (2015) Proteomic analysis of quail calcified eggshell matrix: a comparison to chicken and turkey eggshell proteomes. Proteome Sci 13:22CrossRefGoogle Scholar
  13. 13.
    Mann K (2015) The calcified eggshell matrix proteome of a songbird, the zebra finch (Taeniopygia guttata). Proteome Sci 13:29CrossRefGoogle Scholar
  14. 14.
    Rose M, Hincke MT (2009) Protein constituents of the eggshell: eggshell-specific matrix proteins. CMLS Cell Mol Life Sci 66:2707–2719CrossRefGoogle Scholar
  15. 15.
    Miksik I, Ergang P, Pacha J (2014) Proteomic analysis of chicken eggshell cuticle membrane layer. Anal Bioanal Chem 406:7633–7640CrossRefGoogle Scholar
  16. 16.
    Verdes A, Cho W, Hossain M, Brennan P.L.R., Hanley D, Grim T, Hauber ME, Holford M (2015) Nature’s Palette: Characterization of Shared Pigments in Colorful Avian and Mollusk Shells. PLoS ONE 10:e0143545Google Scholar
  17. 17.
    Sparks NHC (2011) Eggshell pigments: from formation to deposition. Avian Biol Res 4:162–167CrossRefGoogle Scholar
  18. 18.
    Samiullah S, Roberts JR (2013) The location of protoporphyrin in the eggshell of brown-shelled eggs. Poult Sci 92:2783–2788CrossRefGoogle Scholar
  19. 19.
    Maurer G, Portugal SJ, Hauber ME, Mikšík I, Russell D.G.D., Cassey P (2015) First light for avian embryos: eggshell thickness and pigmentation mediate variation in development and UV exposure in wild bird eggs. Funct Ecol 29:209–218CrossRefGoogle Scholar
  20. 20.
    Duval C, Cassey P, Lovell PG, Mikšík I, Reynolds SJ, Spencer KA (2016) Maternal influence on eggshell maculation: implications for cryptic camouflaged eggs. J Ornithol 157:303–310CrossRefGoogle Scholar
  21. 21.
    Mikšík I, Ergang P, Pácha J (2014) Proteomic analysis of chicken eggshell cuticle membrane layer. Anal Bioanal Chem 406:7633–7640CrossRefGoogle Scholar
  22. 22.
    Mikšík I, Sedláková P, Mikuliková K, Eckhardt A (2007) Proteins of insoluble matrix of avian (Gallus gallus) eggshell. Connect Tissue Res 48:1–8CrossRefGoogle Scholar
  23. 23.
    Miksik I, Holan V, Deyl Z (1996) Avian eggshell pigments and their variability. Comp Biochem Physiol B 113:607–612CrossRefGoogle Scholar
  24. 24.
    Brulez K, Mikšík I, Cooney CR, Hauber ME, Lovell PG, Maurer G, Portugal SJ, Russell D, Reynolds SJ, Cassey P (2016) Eggshell pigment composition covaries with phylogeny but not with life history or with nesting ecology traits of British passerines. Ecol Evol 6:1637–1645CrossRefGoogle Scholar
  25. 25.
    Rappsilber J, Mann M, Ishihama Y (2007) Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nat Protoc 2:1896–1906CrossRefGoogle Scholar
  26. 26.
    Kersey PJ, Duarte J, Williams A, Karavidopoulou Y, Birney E, Apweiler R (2004) The international protein index: an integrated database for proteomics experiments. Proteomics 4:1985–1988CrossRefGoogle Scholar
  27. 27.
    Ishihama Y, Oda Y, Tabata T, Sato T, Nagasu T, Rappsilber J, Mann M (2005) Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein. Mol Cell Proteom 4:1265–1272CrossRefGoogle Scholar
  28. 28.
    Mann K, Macek B, Olsen J (2006) Proteomic analysis of the acid-soluble organic matrix of the chicken calcified eggshell layer. Proteomics 6:3801–3810CrossRefGoogle Scholar
  29. 29.
    Cassey P, Thomas GH, Portugal SJ, Maurer G, Hauber ME, Grim T, Lovell PG, Mikšík I (2012) Why are birds’ eggs colourful? Eggshell pigments co-vary with life-history and nesting ecology among British breeding non-passerine birds. Biol J Linn Soc 106:657–672CrossRefGoogle Scholar
  30. 30.
    Rose-Martel M, Du J, Hincke MT (2012) Proteomic analysis provides new insight into the chicken eggshell cuticle. J Proteom 75:2697–2706CrossRefGoogle Scholar
  31. 31.
    Ohmuraya M, Yamamura K (2011) Roles of serine protease inhibitor Kazal type 1 (SPINK1) in pancreatic diseases. Experimental Anim 60:433–444CrossRefGoogle Scholar
  32. 32.
    Miksik I, Holan V, Deyl Z (1994) Quantification and variability of eggshell pigment content. Comp Biochem Physiol A 109:769–772CrossRefGoogle Scholar
  33. 33.
    Lopez-Rull I, Miksik I, Gil D (2008) Egg pigmentation reflects female and egg quality in the spotless starling Sturnus unicolor. Behav Ecol Sociobiol 62:1877–1884CrossRefGoogle Scholar
  34. 34.
    Wang XT, Zhao CJ, Li JY, Xu GY, Lian LS, Wu CX, Deng XM (2009) Comparison of the total amount of eggshell pigments in Dongxiang brown-shelled eggs and Dongxiang blue-shelled eggs. Poult Sci 88:1735–1739CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Institute of Physiology of the Czech Academy of SciencesPrague 4Czech Republic
  2. 2.Institute of Anatomy, First Faculty of MedicineCharles UniversityPrague 4Czech Republic

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