title: Application MALDI TOF on protein identification of vitellogenin in giant grouper (Epinephelus lanceolatus).
abstract: A new proteomics technology has been implemented to study the protein repertoires of developing oocytes of giant grouper (Epinephelus lanceolatus). Knowledge of the chemical composition and physiochemical properties of vitellogenin (Vtg) is necessary to interpret the functional and biological properties attributed during ovulation. Vtg, as a biomarker indicator in sex determination, has been analyzed to determine the sex and maturational status of fish in the absence of the gonad tissue. A male giant grouper was induced by 2 mg/kg of 17ß-estradiol (E2), and blood was sampled at days 0, 1, 3, 5, and 10. SDS-PAGE 1D electrophoresis was used to analyze Vtg protein, and Vtg identification was done with 4800 Plus MALDI TOF/TOF™ mass spectrophotometer (Applied Biosystems/MDS SCIEX, USA). Meanwhile, MS/MS de novo sequencing identified the proteins by matching sequences of tryptic peptides to the known sequences of other species. Vtg was confirmed by MASCOT at 95% significant level, and molecular mass was 187 kDa. Protein resolved on SDS-PAGE as a double band of approximately the same mass as determined with MALDI-TOF. The N-terminal sequences and identification of Vtg were also determined. The potential of using MS methods to understand the structure and function of Vtg is discussed.
authors.affiliation: Fisheries Research Institute (FRI) Tanjong Demong, Besut, Terengganu, 22200, Malaysia, ahmaddaudom@yahoo.com.
authors.forename: Ahmad Daud
authors.initials: AD
authors.lastname: Om
authors.forename: Safiah
authors.initials: S
authors.lastname: Jasmani
authors.forename: Nosrihah
authors.initials: N
authors.lastname: Ismail
authors.forename: S Y
authors.initials: SY
authors.lastname: Yeong
authors.forename: A B
authors.initials: AB
authors.lastname: Abol-Munafi
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004958:Estradiol / Q000494:pharmacology; D008297:Male; D009865:Oocytes / Q000378:metabolism*; D010473:Perciformes / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000662:veterinary; D012732:Sex Determination Analysis / Q000379:methods / Q000662:veterinary*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods / Q000662:veterinary*; D053719:Tandem Mass Spectrometry / Q000662:veterinary; D014819:Vitellogenins / Q000032:analysis* / Q000096:biosynthesis
chemical_list: D014819:Vitellogenins; D004958:Estradiol