Abstract
The protein components of biomineralized structures (matrix proteins) are believed to modulate crystal nucleation and growth, and theraby influence the shape and strength of the final structure. The chicken eggshell contains a complex array of distinct matrix proteins. The most abundant of these was purified to homogeneity by a combination of anionic exchange and hydroxyapatite chromatographies. Antibodies to this protein were raised in rabbit, and utilized for Western blotting and immunohistochemistry. These studies indicated that the 17 kDa antigen (ovocleidin 17, OC-17) is found in the shell gland mucosa, and that only the tubular gland cells were positive. Immunohistochemistry with decalcified shell indicated that OC-17 is uniformly distributed throughout the shell matrix, but concentrated in the mammillary bodies. Our results indicate that this protein is secreted during shell formation and becomes incorporated into this structure. It may therefore play a role in the crystallization process and influence the properties of the resulting eggshell.
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References
Mann S (1988) Molecular recognition in biomineralization. Nature 332:119–124
Lowenstam HA, Weiner S (1989) On biomineralization. Oxford University Press, New York
Addadi L, Weiner S (1992) Control and design principles in biological mineralization. Angew Chem Int Ed (Engl) 31:153–169
Krampitz G, Graser G (1988) Molecular mechanisms of biomineralization in the formation of calcified shells. Ang Chem Inter Ed (Eng) 27:1145–1156
Weiner S, Addadi L (1991) Acidic macromolecules of mineralized tissues: the controllers of crystal formation. Tr Biochem Sci 16:252–256
Marn S, Archibald DD, Didymus JM, Douglas T, Heywood BR, Meldrum FC, Reeves NJ (1993) Crystallization at inorganic-organic interfaces: biominerals and biominetic synthesis. Science 261:1286–1292
Addadi L, Moradian J, Shay E, Maroudas NG, Weiner S (1987) A chemical model for the cooperation of sulfates and carboxylates in calcite crystal nucleation: relevance to biomineralization. Proc Natl Acad Sci USA 84:2732–2736
Mann S, Heywood BR, Rajam S, Birchall JD (1988) Controlled crystallization of CaCO3 under stearic acid monolayers. Nature 334:692–695
Burley RW, Vadehra DV (1989) The avian egg, chemistry and biology. J Wiley & Sons, Toronto
Leach RM (1982) Biochemistry of the organic matrix of the eggshell. Poultry Sci 61:2040–2047
Hincke MT, Bernard A-M, Lee ER, Tsang CPW, Narbaitz R (1992) Soluble protein constituents of the domestic fowl's eggshell. Br Poultr Sci 33:505–516
Hincke MT, Nairn AC (1992) Phosphorylation of elongation factor 2 during Ca2+-mediated secretion from rat parotid acini. Biochem J 282:877–882
Towbin H, Staehelin T, Gordon S (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Staines WA, Meister B, Melander T Nagy JI, Hokfelt T (1988) Three-colour immunofluorescence histochemistry allowing triple labelling with a single section. J Histochem Cytochem 36: 145–151
Singhrao S, Cole G, Henderson WJ, Newman GR (1990) LR white embedding allows a multi-method approach to the analysis of brain tissue from patients with Alzheimer's disease. Histochemistry 22:257–268
Hewick RM, Hunkapiller MW, Hood LE (1981) A gas-liquid solid phase peptide and protein sequenator. J Biol Chem 256: 7990–7997
Matsudaira P (1987) Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membrane. J Biol Chem 262:10035–10038
Parsons AH (1982) Structure of the eggshell. Poultr Sci 61:2013–2021
Hamilton RMG (1986) The microstructure of the hens egg-shell—a short review. Food Microstruct 5:99–110
Simkiss K (1961) Calcium metabolism and avian reproduction. Biol Rev Cambridge Philos Soc 36:321–367
Simkiss K, Tyler C (1957) A histochemical study of the organic matrix of hen egg-shells. Q J Microsc Sci 98:19–28
Robinson DS, King NR (1968) Mucopolysaccharides of an avian egg shell membrane. J R Microsc Soc 88:13–22
Salevsky E, Leach RM (1980) Studies on the organic components of shell gland fluid and the hen's egg shell. Poultr Sci 59:438–4443
Silyn-Roberts H, Sharp RM (1986) Crystal growth and the role of the organic network in eggshell mineralization. Proc Roy Soc (Lond) B227:303–324
Baker JR, Balch DA (1962) A study of the organic material of the hen's egg-shell. Biochem J 82:352–361
Abatangelo G, Daga-Gordini D, Castellani I, Cortivo R (1978) Some observations on the calcium ion binding to the eggshell matrix. Calcif Tissue Res 26:247–252
Eckert J, Glock H, Schade R, Krampitz G, Enbergs H, Petersen J (1986) Synthesis of a precursor polypeptide of egg shell matrix in the liver of laying hen. J Animal Physiol Animal Nutr 56:258–267
Arias JL, Carrino DA, Fernandez MS, Rodriguez JP, Dennis JE, Caplan AI (1992) Partial biochemical and immunochemical characterization of avian eggshell extracellular matrices. Arch Biochem Biophys 298:293–302
Simkiss K, Wilbur KM (1989) Biomineralization, cell biology and mineral deposition. Academic Press, San Diego
Wyburn GM, Johnston HS, Draper MH, Davidson MF (1973) The ultrastructure of the shell-forming region of the oviduct and the development of the shell of Gallus domesticus. Q J Exp Physiol 58:143–151
Creger CR, Phillips H, Scott JT (1976) Formation of an eggshell. Poultr Sci 55:1717–1723
Stemberger BH, Mueller WJ, Leach RM (1977) Microscopic study of the initial stages of eggshell calcification. Poultr Sci 56:537–543
Dieckert JW, Dieckert MC, Creger CR (1989) Calcium reserve assembly: a basic structural unit of the calcium reserve system of the hen egg shell. Poultr Sci 68:1569–1584
Wassermann RH, Smith CA, Smith CM, Brindak ME, Fullmer CS, Krook L, Penniston JT, Kumar R (1991) Immunohistochemical localization of a calcium pump and calbindin-D28k in the oviduct of the laying hen. Histochemistry 96:413–418
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Hincke, M.T., Tsang, C.P.W., Courtney, M. et al. Purification and immunochemistry of a soluble matrix protein of the chicken eggshell (ovocleidin 17). Calcif Tissue Int 56, 578–583 (1995). https://doi.org/10.1007/BF00298593
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DOI: https://doi.org/10.1007/BF00298593