title: On plate graphite supported sample processing for simultaneous lipid and protein identification by matrix assisted laser desorption ionization mass spectrometry.
abstract: The simultaneous identification of lipids and proteins by matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS) after direct on-plate processing of micro-samples supported on colloidal graphite is demonstrated. Taking advantages of large surface area and thermal conductivity, graphite provided an ideal substrate for on-plate proteolysis and lipid extraction. Indeed proteins could be efficiently digested on-plate within 15 min, providing sequence coverages comparable to those obtained by conventional in-solution overnight digestion. Interestingly, detection of hydrophilic phosphorylated peptides could be easily achieved without any further enrichment step. Furthermore, lipids could be simultaneously extracted/identified without any additional treatment/processing step as demonstrated for model complex samples such as milk and egg. The present approach is simple, efficient, of large applicability and offers great promise for protein and lipid identification in very small samples.
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy; Centro Interdipartimentale S.M.A.R.T., Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy; Centro Interdipartimentale, "Laboratorio di ricerca per la diagnostica dei Beni Culturali", Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy. Electronic address: cosimadamiana.calvano@uniba.it.
authors.forename: Cosima Damiana
authors.initials: CD
authors.lastname: Calvano
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy.
authors.forename: Inez Dorothé
authors.initials: ID
authors.lastname: van der Werf
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy; Centro Interdipartimentale, "Laboratorio di ricerca per la diagnostica dei Beni Culturali", Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy.
authors.forename: Luigia
authors.initials: L
authors.lastname: Sabbatini
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy; Centro Interdipartimentale S.M.A.R.T., Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy; Centro Interdipartimentale, "Laboratorio di ricerca per la diagnostica dei Beni Culturali", Università degli Studi di Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Palmisano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006108:Graphite / Q000737:chemistry*; D008055:Lipids / Q000032:analysis* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D059748:Proteolysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
chemical_list: D008055:Lipids; D010455:Peptides; D011506:Proteins; D006108:Graphite; D014357:Trypsin
keywords: Graphite; Lipidomics; MALDI MS; On plate digestion; Phosphopeptides; Proteomics