Abstract:

Objective To study the techniques of the isolation and identification of the proteins in Hela cells. Methods The proteins from Hela cells were extracted and isolated by two-dimensional electrophoresis(2-DE). The digital information of the protein spots was analyzed by a scanner and analysis software. Three spots of proteins in gel with different abundance were digested by an in-gel enzyme and the masses of the generated peptides were measured by matrix assisted laser desorption ionization orthogonal time of flight mass spectrometry(MALDI O-TOF-MS). The data obtained from peptide mass fingerprinting(PMF) were subjected to the protein database search. Results The 2-DE map of the proteins from Hela cells was generated by 2-DE, and the digital information of PMFs for the three protein spots were obtained by MALDI O-TOF-MS. Subsequently, these three protein spots were identified as α-glucosidase, γ-actin and peroxiredoxin 2 respectively through the database searching. Conclusion Peptide mass fingerprinting(PMF) is an applicable technique for the identification of proteins of Hela cells in combination with 2-D gel electrophoresis.

Key words: Hela cells, proteome, two-dimensional electrophoresis, MALDI O-TOF-MS, peptide mass fingerprinting