Skip to main content
Log in

Isolation and identification of yolk proteins in Indian major carp,Labeo rohita

  • Published:
Journal of Biosciences Aims and scope Submit manuscript

Abstract

Two yolk proteins (YP1 and YP2) from the ovaries of Indian major carp, Labeo rohita were isolated by gel filtration and partially characterized by the use of hydroxyapatite ultrogel column in conjunction with native PAGE. On native PAGE YP1 gave a single protein band, whereas YP2 of gel filtration revealed the contamination of YP1, which was removed by adsorption chromatography on hydroxyapatite ultrogel and then the YP2 was the purified one as judged by electrophoresis. Both YP1 and YP2 also stained for lipid and contained alkalilabile phosphorus. Therefore, both yolk proteins were lipophosphoprotein. The molecular weights of YP1 and YP2 were 620 kDa and 225 kDa respectively as determined by gel filtration on Sepharose 4B. When YP1 and YP2 were compared in relation to some physicochemical characteristics with yolk proteins of other oviparous vertebrates including fish, they were lipovitellin like.

Antiserum to YP2 crossreacted with YP2 and vitellogenin suggesting that YP2 was the cleaved product of vitellogenin. Anti-YP2 antiserum was not crossreacted with native YP1, whereas reduced and/or denatured YP1 was crossreacted indicating the presence of antigenic determinants in the inner core region of YP1 polypeptide.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ando K 1965 Ultracentrifugal analysis of yolk proteins in rainbow trout egg and their changes during development;Can. J. Biochem. 43 373–379

    Article  PubMed  CAS  Google Scholar 

  • Andrews P 1965 The gel filtration behaviour of proteins related to their molecular weight over a wide range;Biochem. J. 96 595–606

    PubMed  CAS  Google Scholar 

  • Bergink E W and Wallace R A 1974 Precursor-product relationship between amphibian vitellogenin and phosvitin;J. Biol Chem. 249 2897–2903

    PubMed  CAS  Google Scholar 

  • Bhakta M 1990Purification and characterization of yolk precursor protein in Indian major carp, Labeo rohita, Ph.D thesis, Visva-Bharati University, Santiniketan

    Google Scholar 

  • Burley R W and Cook W H 1961 Isolation and composition of avian egg yolk granules and their constituent α-andβ-lipovitellins;Can. J. Biochem. Physiol. 391 1295–1307

    Google Scholar 

  • Byrne B M, Gruber M and Ab G 1989 The evolution of egg yolk proteins;Prog. Biophys. Mol. Biol. 53 33–69

    Article  PubMed  CAS  Google Scholar 

  • Campbell A. M and Idler D R 1980 Characterization of an estradiol-induced proteins from rainbow trout serum as vitellogenin by the composition and radioimmunological crossreactivity to ovarian yolk fractions:Biol. Report. 22 605–618

    CAS  Google Scholar 

  • Carnevali O, Mosconi G, Roncarati A, Belvedere P, Romano M and Limatola E 1992 Changes in the electrophoretic pattern of yolk proteins during vitellogenesis in the gilthead sea bream,Sparus aurata L;Comp. Biochem. Physiol. B103 955–962

    Google Scholar 

  • Cook W H and Martin W G 1969 Egg lipoproteins; inStructural and functional aspects of lipoproteins in living systems (eds) E Tris and A M Scanu (New York: Academic press) pp 579–615

    Google Scholar 

  • Davis B J 1964 Disc electrophoresis II. Method and application to human serum proteins:Ann. N. Y. Acad. Sci. 121 404–427

    Article  PubMed  CAS  Google Scholar 

  • de Vlaming V L, Wiley H S, Delahunty G and Wallace R A 1980 Goldfish(Carassius auratus) vitelogenin: Induction, isolation, properties and relationship to yolk proteins;Comp. Biochem. Phsiol. B67 613–623

    Article  Google Scholar 

  • Hara A 1987 Studies on female-specific serum protein (vitellogenin) and egg yolk proteins in teleosts: Immunochemical, physicochemical and structural studies;Mem. Fac. Fish. Hokkaido Univ. 34 1–59

    Google Scholar 

  • Hara A and Hirai H 1978 Comparative studies on immunochemical properties of female specific serum protein and egg yolk proteins in rainbow trout(Salmo gairdneri), Comp. Biochem. Physiol. B59 339–343

    Google Scholar 

  • Hara A, Matsubara T, Sancyashi M and Takano K 1984 vitellogenin and its derivatives in egg yolk proteins of white-spotted char(Salivelinus leucomaenis):Bull. Fac. Fish. Hokkaido Univ. 35 144–153

    Google Scholar 

  • Hara A, Sullivan A. V and Dickhoff W W 1993 Isolation and some characterization of vitellogenin and its related egg yolk proteins from Coho Salmon(Oncorhynchus kisutch);Zool. Sci. 10 245–256

    CAS  Google Scholar 

  • Jared D W and Wallace R A 1968 Comparative chromatography of the yolk proteins of teleosts;Comp. Biochem. Physiol. 24 437–443

    Article  PubMed  CAS  Google Scholar 

  • Kishida M and Specker J L 1993 Vitellogenin in tilapia(Oreochromis mossambicus); induction of two forms by estradiol, quantification in the plasma and characterization in oocyte extract;Fish physiol. Biochem. 12 171–182

    Article  CAS  Google Scholar 

  • Kishida M, Anderson T R and Specker J L 1992 Induction by β-estradiol of vitellogenin in stripped bass(Morone saxatilis): Characterization and quantification in plasma and mucus;Gen. Comp. Endocrinol. 88 29–39

    Article  PubMed  CAS  Google Scholar 

  • Markert J R and Vanstone WE 1971 Egg proteins of Coho Salmon(Orcorhynchus kisutch); Chromatographic separation and molecular weight of the major protein in the high density fraction and their presence in Salmon plasma;J. Fish. Res. Board Can. 28 1853–1856

    CAS  Google Scholar 

  • McCollum K, Gregory D, Williams B and Taborsky G 1986 Phosvitin isolation from fish eggs: methodological improvements including specific phosvitin precipitation with ferric ion;Comp. Biochem. Physiol. B84 151–157

    Google Scholar 

  • Mommsen T P and Walsh P J 1988 Vitellogenesis and oocyte assembly; inFish physiology (eds) W S Hoar and D J Randell (New York: Academic press) vol. 11A, pp 347–406

    Google Scholar 

  • Naglar J J, Idler D R and So Y P 1992 A comparison of tritiated vitellogenins prepared byin vivo andin vitro techniques for studies of ovarian uptake in fish;J. Exp. Zool. 264 107–112

    Article  Google Scholar 

  • Nath P and Sundararaj B I 1981 Isolation and identification of female specific serum lipophosphoprotein (Vitellogenin) in the catfish,Heteropnettstes fossilis (Bloch);Gen. Comp. Endocrinol. 43 181–190

    Google Scholar 

  • Nath P, Bhakta M and Mitra K 1992 Demonstration of two forms of Vitellogenin in serum of estradiol-17 β-treated Indian major carp,Labeo rohita;Indian. J. Exp. Biol. 30 464–469

    PubMed  CAS  Google Scholar 

  • Ng T B and Idler D R 1983 Yolk formation and differentiation in teleost fishes; inThe Fish physiology (eds) W S Hoar, D J Randell and E M Donaldson (New York: Academic Press) pp 373–404

    Google Scholar 

  • Ouchterlony O 1953 Antigen-antibody reaction in gels. IV. Types of reaction in coordinated systems of diffusion;Acta. Pathol. Microbiol. Scand. 32 231–240

    CAS  Google Scholar 

  • Plack P A, Pritchard D J and Frazer N A 1971 Egg proteins in cod serum. Natural occurance and induction by injections of estradiol-3-Benzoate;Biochem. J. 121 847–856

    PubMed  CAS  Google Scholar 

  • Prat J P, Lamy J N and Weill J D 1969 Coloration des lipoproteines apres electrophorese in gel de polyacrylamide;Bull. Soc. Chico. Biol. 51 1367

    CAS  Google Scholar 

  • Radomski M W, Wallace R A and Cook W H 1963 Comparative chromatographic and dissociative behaviour of avian and amphibian lipovitellin;Biochem. Biophys. Acta. 70 600–602

    Article  PubMed  CAS  Google Scholar 

  • Shigeura H T and Haschemeyer A E V 1984 Purification of egg yolk proteins from the Antarctic fish,Chaenocephalus aceratus: Comp. Biochem. Physiol. B80 935–939

    Google Scholar 

  • Specker J L and Sullivan C V 1994 Vitellogenesis in fishes; Status and perspectives; inPerspectives in comparative endocrinology (eds) K G Davey R E Peter and S S Tobe (National Research Council of Canada) pp 304-315

  • Tao Y, Hara A, Hodson R G, Woods L C III and Sullivan A. V 1993 Purification, characterization and immunoassay of stripped bass(Morone saxatilis) vitellogenin;Fish. Physiol. Biochem. 12 31–46

    Article  CAS  Google Scholar 

  • Tyler A. R, Sumpter J P and Bromage N R 1988In vivo ovarian uptake and processing of vitellogenin in the rainbow trout,Salmo gairdneri;J. Exp. Zool. 246 171–179

    Article  Google Scholar 

  • Wallace R A 1985 Vitellogenesis and oocyte growth in nonmammalian Vertebrates; inDevelopmental biology (ed.) L W Browder (New York: Plenum) Vol. 1, pp 127–177

    Google Scholar 

  • Wallace R A and Begovac P C 1985 Phosvitins in Fundulus oocytes and eggs: Preliminary chromatographic and electrophoretic analysis together with biological considerations;J. Biol. Chem. 260 11268–11274

    PubMed  CAS  Google Scholar 

  • Wallace R A and Morgan J P 1986 Chromatographic resolution of chicken phosvitin: Multiple macromolecular species in a classic vitellogenin-derived phosphoprotein;Biochem. J. 240 871–878

    PubMed  CAS  Google Scholar 

  • Wallace R A and Selman K 1990 Ultrastructural aspects of oogenesis and oocyte growth in fish and amphibians;J. Electron Microsc. 16 175–201

    Article  CAS  Google Scholar 

  • Wallace R A, Jared D W and Eisen A Z 1966 A general method for the isolation and purification of phosvitin from vertebrate eggs;Can. J. Biochem. 44 1647–1655

    CAS  Google Scholar 

  • Wiley H S and Wallace R A 1981 The structure of vitellogenin. Multiple vitellogenins inXenopus laevis give rise to multiple forms of the yolk proteins;J. Biol. Chem. 256 8626–8634

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to P. Nath.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bhakta, M., Nath, P. Isolation and identification of yolk proteins in Indian major carp,Labeo rohita . J Biosci 21, 711–722 (1996). https://doi.org/10.1007/BF02703147

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02703147

Keywords

Navigation