Phosphorylated membrane proteome analysis of human neuroblastoma SH-SY5Y cell line

Chunyan Hou, Junfeng Ma, Dingyin Tao, Lihua Zhang, Zhen Liang, Yukui Zhang

Research output: Contribution to journalArticle

Abstract

A strategy with the combination of multiprotease digestion and the selective enrichment of phosphopeptides by silica hybrid monolith based immobilized Ti 4+affinity chromatography (Ti 4+-IMAC) was proposed, and applied in the global profiling of phosphorylated membrane proteome of neuroblastoma SH-SY5Y cells. The fraction of membrane proteins was extracted by ultra speed centrifuge, followed by washing with 1 mol/L sodium chloride and 0.1 mol/L sodium carbonate. For digestion, chymotrypsin and pepsin with broader specificity were used as complementary enzymes to trypsin. The phosphopeptides were then selectively enriched by monolithic Ti 4+-IMAC column, and analyzed by nanoflow high performance liquid chromatography and mass spectrometry. A total of 43 phosphoproteins were identified, among which 14 proteins were located on the membrane. All these results demonstrated that the proposed strategy might be promising to promote the in-depth study of neuroblastoma and discover the candidate biomarkers.

Original languageEnglish (US)
Pages (from-to)851-856
Number of pages6
JournalChinese Journal of Chromatography (Se Pu)
Volume29
Issue number9
DOIs
StatePublished - Sep 28 2011

Keywords

  • Human neuroblastoma
  • Immobilized Ti affinity chromatography
  • Membrane protein
  • Multi-enzyme digestion
  • Organic-inorganic hybrid silica monolith
  • Phosphopeptide enrichment

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Chemical Engineering(all)
  • Organic Chemistry
  • Electrochemistry

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