Analysis of nickel-binding peptides in a human hepidermoid cancer cell line by Ni-NTA affinity chromatography and mass spectrometry.
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.
Lamberti
Annalisa
A
Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli Federico II, Via S. Pansini 5, I-80131, Napoli, Italy.
Sanges
Carmen
C
Longo
Olimpia
O
Chambery
Angela
A
Di Maro
Antimo
A
Parente
Augusto
A
Masullo
Mariorosario
M
Arcari
Paolo
P
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D006801:Humans; D008969:Molecular Sequence Data; D009532:Nickel / Q000378:metabolism*; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry*; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000378:metabolism
D000199:Actins; D009942:Organometallic Compounds; D010455:Peptides; C088321:nickel nitrilotriacetic acid; C022838:nickel chloride; D009532:Nickel; D014357:Trypsin; D009571:Nitrilotriacetic Acid