Metal imaging in non-denaturating 2D electrophoresis gels by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for the detection of metalloproteins.

Ryszard Lobinski (1) , J. Sabine. Becker

1 IPREM - Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux

Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was developed as a powerful anal. technique for metal imaging of 2D gels for the detection of metalloproteins in rat kidney after electrophoretic sepn. Protein complexes, extd. with water, were sepd. in their native state in the first and second dimension by blue native gel electrophoresis (BN-PAGE). Essential and toxic metals, such as zinc, copper, iron, manganese and lead, were monitored by LA-ICP-MS after gel ablation by a focused laser beam in a way that the total surface of a selected fragment of the gel was totally ablated. The metal distribution of this part of the gel was then constructed by plotting the metal (isotope) signal intensity as a function of the x,y (isoelec. point, mol. mass) coordinates of the gel. The proteins at locations rich in metals were cut out, digested with trypsin and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

Mots clés: metal imaging electrophoresis gel LA ICP MS metalloprotein

Domaines: Chimie analytique

Ryszard Lobinski, J. Sabine. Becker. Metal imaging in non-denaturating 2D electrophoresis gels by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for the detection of metalloproteins.. Metallomics, 2009, 1, pp.312-316. ⟨10.1039/b822974k⟩. ⟨hal-01560748⟩