NanoLC-MS coupling of liquid microjunction microextraction for on-tissue proteomic analysis

Biochim Biophys Acta Proteins Proteom. 2017 Jul;1865(7):891-900. doi: 10.1016/j.bbapap.2016.11.002. Epub 2016 Nov 9.

Abstract

Mass spectrometry (MS)-based microproteomics on localized regions of tissue sections was achieved by direct coupling of liquid microjunction microextraction with a nanoscale liquid chromatography-tandem MS, resulting in the identification of >500 protein groups from a region as small as 250μm in diameter representing only a few hundred of cells. The method was applied on the examination of benign and tumor regions initially defined by imaging mass spectrometry (IMS) analysis of a consecutive high grade serous ovarian tumor tissue section. Results identified the higher abundance of eukaryotic translation initiation factors eIF4A, its isoform eIF4A2, and eIF5A and its isoform eIF5A2, and lower abundance of actin-binding proteins OBSCN, TAGLN and CNN3 on tumor regions, concomitant with previous findings. This demonstrates the use of the method for downstream characterization of distinct regions identified by IMS. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann.

Keywords: Imaging mass spectrometry; Liquid microjunction; Microextraction; Microproteomics; Ovarian cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chemical Fractionation / methods
  • Chromatography, Liquid / methods
  • Eukaryotic Initiation Factor-4A / metabolism
  • Eukaryotic Translation Initiation Factor 5A
  • Humans
  • Microfilament Proteins / metabolism
  • Neoplasms / diagnosis
  • Neoplasms / metabolism
  • Neoplasms / pathology
  • Peptide Initiation Factors / metabolism
  • Proteins / metabolism*
  • Proteome / metabolism*
  • Proteomics / methods
  • RNA-Binding Proteins / metabolism
  • Tandem Mass Spectrometry / methods

Substances

  • Microfilament Proteins
  • Peptide Initiation Factors
  • Proteins
  • Proteome
  • RNA-Binding Proteins
  • Eukaryotic Initiation Factor-4A