Assessing matrix assisted laser desorption/ ionization-time of flight-mass spectrometry as a means of rapid embryo protein identification in rice

• Published in: Electrophoresis 2003-04, Vol.24 (7-8), p.1319-1329
• Main Authors: Fukuda, Masafumi, Islam, Nazrul, Woo, Sun-Hee, Yamagishi, Azumi, Takaoka, Motoko, Hirano, Hisashi
• Format: Article
• Language: English
• Subjects: Databases, Protein; Electrophoresis, Gel, Two-Dimensional; Oryza - chemistry; Peptide mass fingerprinting; Plant Proteins - analysis; Protein identification; Proteomics - methods; Rice (Oryza sativa L.); Seeds - chemistry; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Time Factors; Two-dimensional electrophoresis
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.200390168

Rice embryo proteins were separated by two‐dimensional gel electrophoresis (2‐DE). A total of 105 spots were digested with trypsin and the resultant peptides were analyzed by matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS). Raw mass spectra were fully‐automatically processed and searched with selected monoisotopic masses against SWISS‐PROT/TrEMBL and NCBInr databases. High quality mass spectra were obtained from 53 spots, of which 36 spots were identified including 29 not registered in databases. Fifty percent of the rice embryo proteins resolved in 2‐DE could not be identified, indicating more efficient sample preparation techniques need to be developed in the future. At least four to five matching peptides were found to be essential for unambiguous identification of rice embryo proteins; peptide matching of less than four lead to ambiguous results. The suitability of peptide mass fingerprinting method as a means of rapid embryo protein identification in rice was discussed.