Identification of Mas Partner Proteins Based on the SPR-MS Coupling Technique
Objective To optimize a surface plasmon resonance(SPR)-mass spectrometry(MS)coupling technique for the identification of Mas partner proteins in cell lysates.Methods The recombinant proteins GST-Mas-CT and GST were immobilized at a high level on 4 channels of 2 CM5 chips,respectively;cell lysates(total protein concentration:0.5 mg·mL-1)were injected over the sensor surface;the recovered binding proteins were prepared using filter-assisted sample preparation(FASP)and identified using MS method;the differential interacting proteins were analyzed with Venn Diagram,and the GO and KEGG analyses were performed by DAVID database.Results A total of 785 Mas-CT differential interacting proteins were found in the recovered proteins;bioinformatics analysis showed that Mas partner proteins were primarily involved in biological processes or pathways,such as immune response,calcium ion response,infection,and platelet activation.Conclusions The optimized SPR-MS coupling technique can be utilized for fishing and identifying partner proteins of target proteins from complex systems such as cell lysates.
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