Induction and partial characterization of California halibut ( Paralichthys californicus) vitellogenin

The egg yolk precursor protein, vitellogenin (Vg), was isolated by size exclusion and ion exchange chromatography from plasma of California halibut ( Paralichthys californicus) treated with estrogen. MALDI TOF mass spectrometry (MS) analysis resulted in a molecular mass of 188 kDa. MS/MS de novo seq...

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Bibliographic Details
Published in:Comparative biochemistry and physiology. Part A, Molecular & integrative physiology Molecular & integrative physiology, 2007-02, Vol.146 (2), p.200-207
Main Authors: Palumbo, A.J., Linares-Casenave, J., Jewell, W., Doroshov, S.I., Tjeerdema, R.S.
Format: Article
Language:English
Subjects:
Amino acid composition
Amino Acid Sequence
Animals
Blotting, Western
Brackish
California halibut
Electrophoresis, Polyacrylamide Gel
Estradiol - pharmacology
Flounder - physiology
Freshwater
Male
Marine
Mass spectrometry
Molecular Sequence Data
MS/MS de novo sequencing
Multiple forms
Oryzias latipes
Paralichthys californicus
Scophthalmus maximus
SDS-PAGE
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Tandem Mass Spectrometry
Vitellogenin
Vitellogenins - biosynthesis
Vitellogenins - chemistry
Western blot
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Summary:The egg yolk precursor protein, vitellogenin (Vg), was isolated by size exclusion and ion exchange chromatography from plasma of California halibut ( Paralichthys californicus) treated with estrogen. MALDI TOF mass spectrometry (MS) analysis resulted in a molecular mass of 188 kDa. MS/MS de novo sequencing identified the protein as Vg by matching sequences of tryptic peptides to the known sequences of several other species. Matches were also made to two different forms of Vg in haddock, medaka, and mummichog, providing evidence that California halibut has more than one form of Vg. Native PAGE and Western blot with an antibody to turbot ( Scophthalmus maximus) Vg confirmed the identity of the protein. Protein resolved on the SDS PAGE as a double band of approximately the same mass as determined with MALDI TOF, and two lower mass bands that were also immunoreactive. MALDI TOF and MS/MS de novo sequencing were useful for determining the molecular mass, identification, and exploring the multiplicity of Vg. The potential of using other MS methods to understand the structure and function of Vg is discussed.
ISSN:1095-6433
1531-4332
DOI:10.1016/j.cbpa.2006.10.023