title: Metal imaging in non-denaturating 2D electrophoresis gels by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for the detection of metalloproteins.
abstract: Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was developed as a powerful analytical technique for metal imaging of 2D gels for the detection of metalloproteins in rat kidney after electrophoretic separation. Protein complexes, extracted with water, were separated 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 (isoelectric point, molecular 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).
authors.affiliation: Laboratoire de Chimie Analytique Bio-inorganique et Environnement, UMR 5254, CNRS, Hélioparc, 64053 Pau, France. susanne.becker@sandoz.com
authors.forename: J Susanne
authors.initials: JS
authors.lastname: Becker
authors.forename: Ryszard
authors.initials: R
authors.lastname: Lobinski
authors.forename: J Sabine
authors.initials: JS
authors.lastname: Becker
mesh_terms: D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D007668:Kidney / Q000737:chemistry; D007834:Lasers; D008667:Metalloproteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D019216:Metals, Heavy / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism; D014867:Water
chemical_list: D008667:Metalloproteins; D019216:Metals, Heavy; D010446:Peptide Fragments; D014867:Water; D014357:Trypsin