Analysis of Nickel-Binding Peptides in a Human Hepidermoid Cancer Cell Line by Ni-NTA Affinity Chromatography and Mass Spectrometry

Title: Analysis of Nickel-Binding Peptides in a Human Hepidermoid Cancer Cell Line by Ni-NTA Affinity Chromatography and Mass Spectrometry

Volume: 15 Issue: 10

Author(s): Annalisa Lamberti, Carmen Sanges, Olimpia Longo, Angela Chambery, Antimo Di Maro, Augusto Parente, Mariorosario Masullo and Paolo Arcari

Affiliation:

Keywords: Affinity chromatography, nickel-binding protein, tandem mass spectrometry, actin, tubulin, elongation factor 1A

Abstract: To elucidate whether eukaryotic elongation factor 1A (eEF-1A) in a human hepidermoid cancer cell line (H1355) belonged to the family of the Ni-interacting protein, we analyzed the sequence of peptides obtained by on-Ni- NTA-agarose tryptic digestion of proteins from H1355 cell extract. LC/MS analysis showed the presence of several peptides mainly from abundant cellular proteins corresponding to eEF-1A, tubulin and actin. The results indicated that F-actin strongly binds to Ni-NTA-agarose whereas the other proteins are indirectly bound to the resin because of the formation of a protein-protein complex with actin.

Cite this article as:

Lamberti Annalisa, Sanges Carmen, Longo Olimpia, Chambery Angela, Maro Di Antimo, Parente Augusto, Masullo Mariorosario and Arcari Paolo, Analysis of Nickel-Binding Peptides in a Human Hepidermoid Cancer Cell Line by Ni-NTA Affinity Chromatography and Mass Spectrometry, Protein & Peptide Letters 2008; 15 (10) . https://dx.doi.org/10.2174/092986608786071157