PMID: PMC6399673-1-2

 

    Legend: Gene, Sites

Title : MS-Based Identification of MGL-Binding Proteins from JurkatCells

Abstract :

1. Next, we performed three biologically independent pull-downexperiments with MGL-Fc and analyzed the bound proteins by LC–MS/MSfollowing trypsin digestion
2. As a negative control, pull-down assaysin the presence of EDTA were performed
3. Altogether, these experimentsresulted in the identification of 775 proteins (data not shown), ofwhich 540 were identified in at least two experiments
4. To filter for proteins specifically binding to MGL , we selected proteins that werenot observed in the negative controls and at least two times in thepull-downs with MGL or proteins that were found in all three pull-downswith MGL and at most once in a negative control
5. This resulted ina list of 85 proteins (Table S1)
6. The candidateMGL-binding proteins included intracellular, plasma membrane, andpredicted secreted proteins
7. Because we used total cell lysates forthe pull-down assays, some of these proteins may be physiologicallyless relevant
8. To filter for cell-surface proteins , which may be expectedto be in direct contact with MGL , we next selected only those proteinsthat were annotated as cell-surface proteins in the UniProtKB database ,resulting in a final list of 17 MGL-binding cell-surface proteins ( Table )
9. Importantly,these proteins were not found in the negative control samples (seealso Table S1), making them strong candidateMGL-binding proteins
10. As expected, CD45 ( PTPRC ) and CD43 ( SPN ) werethe two main MGL-binding proteins identified based on the total numberof peptide-spectrum matches (summed for the three biological replicates)for these two proteins , respectively

Output (sent_index, trigger, protein, sugar, site):

Output(Part-Of) (sent_index, protein, site):

*Output_Site_Fusion* (sent_index, protein, sugar, site):

 

Protein	NCBI ID	SENTENCE INDEX