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Updated: May 26, 2026
Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line
Published on: February 17, 2016
[Phosphorylated membrane proteome analysis of human neuroblastoma SH-SY5Y cell line]
Chunyan Hou1, Junfeng Ma, Dingyin Tao
1CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physic Chinese Academy of Sciences, National Chromatographic R. & A. Center, Dalian 116023, China.
Abstract:
A strategy with the combination of multiprotease digestion and the selective enrichment of phosphopeptides by silica hybrid monolith based immobilized Ti4+ affinity chromatography (Ti4+ -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 Ti4+ -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.
