Table 4.

SILAC analysis of K562 cells to quantify phosphorylation site responsiveness to imatinib


PSDB ID

pTyr site

Peptide

Control/treated
Peptides that are less abundant following imatinib treatment    
    Adaptor/scaffold     
        Abi-1   213   TLEPVKPPTVPNDyMTSPAR   > 2  
        CD2AP   548   DTCYSPKPSVyLSTPSSASK   > 2  
        Gab1   406   DASSQDCyDIPR   > 4  
        Shc   427   ELFDDPSyVNVQNLDK   > 10  
    Protein kinase     
        Abl   115   NGQGWVPSNyITPVNSLEK   > 10  
        Abl   393   LMTGDTyTAHAGAK   > 20  
        Abl   393   VADFGLSRLMTGDTyTAHAGAK   > 10  
        Bcr   177   GHGQPGADAEKPFyVNVEFHHER   > 2  
        Bcr   177   KGHGQPGADAEKPFyVNVEFHHER   > 10  
        Bcr   246   SSESSCGVDGDyEDAELNPR   > 2  
        Bcr   591   LASQLGVyR   > 5  
        Bcr   644   NSLETLLyKPVDR   > 5  
        ERK2   187   VADPDHDHTGFLTEyVATR   > 2  
    Lipid phosphatase     
        SHIP-2   986   NSFNNPAYyVLEGVPHQLLPPEPPSPAR   > 20  
        SHIP-2   1135   TLSEVDyAPAGPAR   > 10  
    Transcription factor     
        STAT5A   694   AVDGyVKPQIK   > 20  
    Other     
        LIM   142   YTEFyHVPTHSDASK   > 2  
        ZO2   1118   IEIAQKHPDIyAVPIK   > 2  
Peptide intensities remaining unchanged following imatinib treatment    
    Protein kinase     
        Cdc2   15   IEKIGEGTyGVVYK   1.10* 
        Cdc2   15   IGEGTyGVVYK   1.04* 
        Cdc2   15   IGEGTyGVVYKGR   1.04* 
        DYRK1A   321   IYQyIQSR   1.04* 
        GSK3-β   216   GEPNVSyICSR   1.22* 
        HIPK1   352   AVCSTyLQSR   1.03* 
        p-38-α   182   HTDDEMTGyVATR   1.61  
        PRP4   849   LCDFGSASHVADNDITPyLVSR   1.14* 
    Other     
        eEF1A-1   29   STTTGHLIyK   1.33  
Peptides without definitive quantification    
    Adaptor/scaffold     
        Cbl   674   IKPSSSANAIySLAARPLPVPK   ND  
    Other     
        Calmodulin
 
100
 
VFDKDGNGyISAAELR
 
ND
 

PSDB ID

pTyr site

Peptide

Control/treated
Peptides that are less abundant following imatinib treatment    
    Adaptor/scaffold     
        Abi-1   213   TLEPVKPPTVPNDyMTSPAR   > 2  
        CD2AP   548   DTCYSPKPSVyLSTPSSASK   > 2  
        Gab1   406   DASSQDCyDIPR   > 4  
        Shc   427   ELFDDPSyVNVQNLDK   > 10  
    Protein kinase     
        Abl   115   NGQGWVPSNyITPVNSLEK   > 10  
        Abl   393   LMTGDTyTAHAGAK   > 20  
        Abl   393   VADFGLSRLMTGDTyTAHAGAK   > 10  
        Bcr   177   GHGQPGADAEKPFyVNVEFHHER   > 2  
        Bcr   177   KGHGQPGADAEKPFyVNVEFHHER   > 10  
        Bcr   246   SSESSCGVDGDyEDAELNPR   > 2  
        Bcr   591   LASQLGVyR   > 5  
        Bcr   644   NSLETLLyKPVDR   > 5  
        ERK2   187   VADPDHDHTGFLTEyVATR   > 2  
    Lipid phosphatase     
        SHIP-2   986   NSFNNPAYyVLEGVPHQLLPPEPPSPAR   > 20  
        SHIP-2   1135   TLSEVDyAPAGPAR   > 10  
    Transcription factor     
        STAT5A   694   AVDGyVKPQIK   > 20  
    Other     
        LIM   142   YTEFyHVPTHSDASK   > 2  
        ZO2   1118   IEIAQKHPDIyAVPIK   > 2  
Peptide intensities remaining unchanged following imatinib treatment    
    Protein kinase     
        Cdc2   15   IEKIGEGTyGVVYK   1.10* 
        Cdc2   15   IGEGTyGVVYK   1.04* 
        Cdc2   15   IGEGTyGVVYKGR   1.04* 
        DYRK1A   321   IYQyIQSR   1.04* 
        GSK3-β   216   GEPNVSyICSR   1.22* 
        HIPK1   352   AVCSTyLQSR   1.03* 
        p-38-α   182   HTDDEMTGyVATR   1.61  
        PRP4   849   LCDFGSASHVADNDITPyLVSR   1.14* 
    Other     
        eEF1A-1   29   STTTGHLIyK   1.33  
Peptides without definitive quantification    
    Adaptor/scaffold     
        Cbl   674   IKPSSSANAIySLAARPLPVPK   ND  
    Other     
        Calmodulin
 
100
 
VFDKDGNGyISAAELR
 
ND
 

ND indicates not determined.

*

Cdc2, DYRK1A, GSK3-beta, HIPK1 and PRP4 were not expected to be drug sensitive from the signature data, and can be viewed as a representative ratio of control/treated.

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