On plate graphite supported sample processing for simultaneous lipid and protein identification by matrix assisted laser desorption ionization mass spectrometry

Talanta. 2015 May:137:161-6. doi: 10.1016/j.talanta.2015.01.030. Epub 2015 Feb 2.

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.

Keywords: Graphite; Lipidomics; MALDI MS; On plate digestion; Phosphopeptides; Proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Graphite / chemistry*
  • Lipids / analysis*
  • Lipids / isolation & purification
  • Molecular Sequence Data
  • Peptides / analysis
  • Peptides / chemistry
  • Proteins / analysis*
  • Proteins / chemistry
  • Proteins / isolation & purification
  • Proteins / metabolism
  • Proteolysis
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
  • Time Factors
  • Trypsin / metabolism

Substances

  • Lipids
  • Peptides
  • Proteins
  • Graphite
  • Trypsin