Summary
Analysis of yolk proteins of the silkworm,Bombyx mori, by SDS-polyacrylamide gel electrophoresis and immunoblotting showed that there was a developmental change in subunit composition of egg-specific protein; egg-specific protein consisting of 72 kDa subunits alone (premature form) was found in vitellogenic follicles, whereas the protein in mature eggs was composed of 72 kDa and 64 kDa subunits (mature form). The premature form of egg-specific protein was purified from young ovaries to homogeneity using a high performance liquid chromatography system. The purified protein had an apparent molecular mass of 225 kDa which could not be distinguished from that of the mature form. By circular dichroism analysis, both egg-specific proteins were estimated to have about 30% α-helix and 20% β-sheet, but the mature form showed a relatively rigid conformation in the aromatic region. The premature egg-specific protein purified from vitellogenic ovaries, consisted of three 72 kDa subunits, whereas mature egg-specific protein was composed of two 72 kDa subunits and one 64 kDa subunit. All of these subunits showed the same immunoreactivity towards antiserum raised against the mature form. An identical NH2-terminal amino acid sequence was found in both 72 kDa polypeptides and 64 kDa polypeptide for the initial 10 amino acids.
Similar content being viewed by others
Abbreviations
- SDS :
-
sodium dodecyl sulfate
- PMSF :
-
phenylmethylsulfonyl fluoride
- PAGE :
-
polyacrylamide gel electrophoresis
- HPLC :
-
high performance liquid chromatography
- ESP :
-
egg-specific protein
- Vtn :
-
vitellin
References
Baert JC (1985) Multiple forms of vitellin in young oocytes ofPerinereis cultrifera (polychaete annelid): Occurrence and relation to vitellin maturation in the oocyte. Comp Biochem Physiol 81B:851–856
Bast RE, Telfer WH (1976) Follicle cell protein synthesis and its contribution to the yolk of theCecropia moth oocyte. Dev Biol 52:83–97
Bergink EW, Wallace RA (1974) Precursor-product relationship between amphibian vitellogenin and the yolk proteins, lipovitellin and phosvitin. J Biol Chem 249:2897–2903
Brookes VJ, Dejmal RK (1968) Yolk protein: Structural changes during vitellogenesis in the cockroachLeucophaea maderae. Science 160:999–1001
Burnette WN (1981) ‘Western Blotting’: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Analyt Biochem 112:195–203
Chen YS, Yang JT, Martinez HM (1972) Determination of the secondary structures of proteins by circular dichroism and optical rotary dispersion. Biochemistry 22:4120–4131
Davis BJ (1964) Dis electrophoresis. II. Method and application to human serum protein. Ann NY Acad Sci USA 131:404–427
Engelmann F (1979) Insect vitellogenin: identification, biosynthesis and role in vitellogenesis. Adv Insect Physiol 14:49–108
Greenfield N, Fasman GD (1969) Computed circular dichroism spectra for the evaluation of protein conformation. Biochemistry 8:4108–4116
Hagedorn HH, Kunkel JG (1979) Vitellogenin and vitellin in insects. Annu Rev Ent 24:475–505
Indrasith LS, Furusawa T, Shikata M, Yamashita O (1987) Limited degradation of vitellin and egg-specific protein inBombyx eggs during embryogenesis. Insect Biochem 17:539–545
Irie K, Yamashita O (1980) Changes in vitellin and other yolk proteins during embryonic development of the silkworm,Bombyx mori. J Insect Physiol 26:811–817
Irie K, Yamashita O (1983) Egg-specific protein in the silkwormBombyx mori: Purification, properties, localization and titre changes during oogenesis and embryogenesis. Insect Biochem 13:71–80
Koeppe J, Ofengand J (1976) Juvenile hormone-induced biosynthesis of vitellogenin inLeucophaea maderae. Arch Biochem Biophys 173:100–113
Kunkel JG, Nordin JH (1985) Yolk proteins. In: Kerkut GA, Gilbert LI (eds) Comprehensive insect physiology, biochemistry and pharmacology, vol 1. Pergamon Press, Oxford, pp 83–111
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227:680–685
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Ono S, Nagayama H, Shimura K (1975) The occurrence and synthesis of female and egg-specific proteins in the silkworm,Bombyx mori. Insect Biochem 5:313–329
Ouchterlony O (1968) Immunodiffusion and immunoelectrophoresis. In: Weir DM (ed) Handbook of experimental immunology. Blackwell Scientific Publications, Oxford, pp 655–706
Shirk PD, Bean D, Millemann AM, Brookes VJ (1984) Identification, synthesis and characterization of the yolk polypeptides ofPlodia interpunctella. J Exp Zool 232:87–98
Telfer WH, Kulakosky PC (1984) Isolated hemolymph proteins as probes of endocytotic yolk formation. Adv Invertebr Reprod 3:81–86
Wallace RA (1985) Vitellogenesis and oocyte growth in nonmammalian vertebrate. In: Browder LW (ed) Developmental biology, vol 1. Plenum, New York, pp 127–177
Woody RW (1978) Aromatic side-chain contributions to the far ultraviolet circular dichroism of peptides and proteins. Biopolymers 17:1451–1467
Wyatt GR, Pan ML (1978) Insect plasma proteins. Annu Rev Biochem 47:779–817
Yamashita O (1986) Yolk protein system inBombyx eggs: Synthesis and degradation of egg-specific protein. Adv Invertebr Reprod 4:79–84
Yamauchi H, Yoshitake N (1984) Developmental stages of ovarian follicles of the silkworm,Bombyx mori. J Morphol 179:21–31
Zhu J, Indrasith LS, Yamashita O (1986) Characterization of vitellin, egg-specific protein and 30 kDa protein fromBombyx eggs, and their fates during oogenesis and embryogenesis. Biochim Biophys Acta 882:427–436
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Indrasith, L.S., Sasaki, T., Yaginuma, T. et al. The occurrence of a premature form of egg-specific protein in vitellogenic follicles ofBombyx mori . J Comp Physiol B 158, 1–7 (1988). https://doi.org/10.1007/BF00692723
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00692723