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Table 2 Differentially regulated phosphopeptides in SS RBCs in response to MEK1/2 inhibitor U0126

From: Proteomic analysis of ERK1/2-mediated human sickle red blood cell membrane protein phosphorylation

Protein description Modified peptide sequence AA vs SS: Fold Change (p-value) SS vs SS+U0126: Fold change (p-value) SS vs SS+ERK2: Fold change (p-value) SS+U0126 vs SS+U0126+ERK2: Fold change (p-value) AA vs AA+ERK2: fold change (p-value)
60S acidic ribosomal protein P2 KEES*EES*DDDMGFGLFD 4.8 (7.24E-20) −2.1 (3.61E-08) NS NS NS
Adenylyl cyclase-associated protein 1 SGPKPFSAPKPQTS*PSPK NS −4.8 (2.00E-03) NS 5.3 (2.00E-04) NS
Alpha-adducin QKGS*EENLDEAR 2.5 (2.43E-33) −4.1 (2.80E-45) NS 2.4 (1.25E-11) NS
Beta-adducin ETAPEEPGS*PAKS*APAS*PVQSPAK 5.6 (3.72E-28) 1.9 (3.87E-09)* NS NS 2.4 (4.21E-06)*
Beta-adducin ETAPEEPGSPAKS*APAS*PVQSPAK NS 1.8 (1.80E-02)* NS 1.8 (3.00E-02) 1.8 (2.00E-02)*
Beta-adducin TESVTSGPMSPEGSPSKS*PSK NS −1.9 (3.00E-02) NS 1.9 (3.00E-03) NS
Dematin QPLTSPGSVS*PSR 7.9 (8.63E-05) −4.4 (1.00E-03) NS NS NS
E3 ubiquitin-protein ligase UBR4 T*SPADHGGSVGSESGGSAVDSVAGEHSVSGR 3.6 (1.55E-18) −1.8 (1.02E-07) NS NS NS
Eukaryotic translation initiation factor 4B SQS*SDTEQQSPTSGGGK 5.6 (9.74E-06) −2.5 (4.00E-03) NS 2.0 (2.00E-02) NS
facilitated glucose transporter member 1 QGGAS*QSDKTPEELFHPLGADSQV 5.0 (7.45E-10) −3.4 (7.99E-08) NS NS NS
Glycophorin-A KS*PSDVKPLPSPDTDVPLSSVEIENPETS*DQ 7.7 (9.74E-07) 1.9 (2.10E-02) NS NS 2.0 (5.00E-03)
Glycophorin-A KSPSDVKPLPS*PDT*DVPLS*SVEIENPETSDQ 10.2 (1.05E-06) 2.3 (5.00E-03) NS NS 2.1 (8.04E-04)
Glycophorin-A S*PS*DVKPLPSPDTDVPLSSVEIENPETS*DQ 6.3 (9.08E-07) 2.0 (1.60E-02) NS NS 1.9 (3.10E-02)
Glycophorin-A S*PS*DVKPLPSPDTDVPLSSVEIENPETSDQ 6.3 (9.08E-07) −2.0 (8.00E-03) NS NS NS
Glycophorin-A SPSDVKPLPS*PDT*DVPLS*SVEIENPETSDQ 7.4 (8.28E-06) 1.9 (2.90E-02) NS NS 1.9 (6.00E-03)
Glycophorin-A SPSDVKPLPS*PDT*DVPLSSVEIENPETSDQ 5.0 (0.00E+00) −2.0 (2.13E-07) NS NS NS
Glycophorin-A SPSDVKPLPSPDT*DVPLS*SVEIENPETSDQ 5.0 (0.00E+00) −1.8 (4.80E-02) NS NS NS
Glycophorin-A SPSDVKPLPSPDT*DVPLSSVEIENPETSDQ 5.0 (0.00E+00) −2.0 (2.59E-21) NS NS NS
Glycophorin-A SPSDVKPLPSPDTDVPLS*S*VEIENPETSDQ 5.1 (3.49E-06) 2.2 (3.00E-03) NS NS 2.7 (7.97E-05)
Glycophorin-A SPSDVKPLPSPDTDVPLSS*VEIENPETSDQ NS −1.9 (3.30E-07) NS NS NS
Glycophorin-A SPSDVKPLPSPDTDVPLSSVEIENPET*SDQ NS −1.9 (1.82E-13) NS NS NS
Glycophorin-A SPSDVKPLPSPDTDVPLSSVEIENPETS*DQ NS −1.8 (4.00E-03) NS NS NS
Leucine-rich repeats and immunoglobulin-like domains protein 2 T*HPETIIALRGMNVTLTCTAVSSSDSPMST*VWR NS −2.9 (1.27E-23) NS 1.8 (2.65E-05) NS
Leucine-zipper-like transcriptional regulator 1 MAGPGST*GGQIGAAALAGGAR 88.6 (1.70E-02) −6.5 (4.50E-02) NS 6.7 (4.54E-06) NS
Lipin-2 S*GGDETPSQSSDISHVLETETIFTPSSVK 3.0 (1.56E-08) −1.9 (3.37E-04) NS NS NS
Metabotropic glutamate receptor 7 LSHKPSDRPNGEAKT*ELCENVDPNS*PAAK 2.4 (1.61E-15) 1.9 (2.95E-09) NS NS 2.3 (7.98E-15)
Proteasome subunit alpha type-3 ESLKEEDES*DDDNM 2.8 (4.14E-11) −1.8 (5.42E-06) NS NS NS
Protein MICAL-2 VS*S*GIGAAAEVLVNLY*MNDHRPKAQAT*SPDLESMRK NS −4.1 (8.44E-05) NS NS NS
Protein Wnt-16 HERWNCMITAAATTAPMGASPLFGYELS*SGTK −2.2 (5.13E-05) −2.0 (1.90E-02) NS NS NS
Spectrin beta chain, erythrocyte LS*SS*WESLQPEPSHPY 3.5 (4.17E-13) 1.8 (5.56E-05) NS NS 2.0 (9.30E-11)
Spectrin beta chain, erythrocyte QIAERPAEETGPQEEEGETAGEAPVS*HHAATER 2.1 (6.00E-03) −2.4 (5.77E-04) NS NS NS
Transmembrane protein 151B SPPGS*AAGES*AAGGGGGGGGPGVSEELTAAAAAAAADEGPAR NS −2.9 (3.91E-05) NS NS NS
U3 small nucleolar RNA-associated protein 15 VVHS*FDYAAS*ILSLALAHEDETIVVGMTNGILS*VKHR NS 2.9 (7.39E-11) NS 2.2 (6.08E-05) 2.1 (9.39E-14)
Uncharacterized protein LOC388588 DGVS*LGAVS*STEEASR NS 1.8 (9.99E-05) NS NS 2.2 (2.40E-09)
Uncharacterized protein LOC388588 DGVS*LGAVSST*EEASR 2.2 (2.00E-02) −1.8 (4.90E-02) NS NS NS
UV excision repair protein RAD23 homolog A EDKS*PSEESAPTTSPESVSGSVPSSGSSGR 5.0 (6.40E-38) 2.0 (1.96E-14) NS NS 2.2 (4.13E-32)
  1. 36 unique phosphorylated peptides were statistically down-regulated in SS RBCs upon treatment with the MEK1/2 inhibitor U0126 (SS vs SS+U0126). Of these 36, 8 phosphopeptides had their levels increased back to near basal levels upon exogenous addition of active ERK2 (SS+U0126 vs SS+U0126+ERK2). Thirteen of these phosphorylated peptides were also observed to be increased in AA RBCs upon exogenous addition of active ERK2 (AA vs AA+ERK2), suggesting specificity of downstream ERK activity. p-values < 0.05 indicate the statistically significant variances associated with differences within triplicates for that particular comparison. Biological variances associated with differences within groups are not included in these p-values. (*) indicates previously reported values[18]. NS, which refers to non-significant, indicates that a statistically significant change (based on 1.75 fold-changes and p-value < 0.05) was not measured.