Table 1.

Integrative list of deformation-induced phosphorylations in the reticulocyte

Accession no.Gene symbolProtein nameSequenceCountSite
E9PIJ1 AMPD2 AMP deaminase 2 TDSDSDLQLYK 7* S190/S197 
A0A0U1RQT1 ACAP2 Arf-GAP with coiled-coil, ANK repeat, and PH domain-containing protein 2 SSPSTGSLDSGNESK 7* S384 
B7Z3I9 ALAD Δ-aminolevulinic acid dehydratase SSPAFGDR S215 
P49840 GSK3A Glycogen synthase kinase-3 α GEPNVSYICSR 7* Y279 
P49840 GSK3A Glycogen synthase kinase-3 α TSSFAEPGGGGGGGGGGPGGSASGPGGTGGGK 7* S21/S39 
E9PK89 RAB3IL1 Guanine nucleotide exchange factor for Rab-3A TLVITSTPASPNR S168 
B4DMA2 HSP90AB1 Heat shock protein HSP 90-β IEDVGSDEEDDSGKDK 7* S255 
Q9H1E3 NUCKS1 Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 VVDYSQFQESDDADEDYGR 7* S19 
S4R2Y4 CBX3 Chromobox protein homolog 3 SLSDSESDDSK S93 
Q08495 DMTN Dematin DSSVPGSPSSIVAK S26 
Q9UG54 MAP3K7 Mitogen-activated protein kinase kinase kinase 7 SIQDLTVTGTEPGQVSSR S439 
P61006 RAB8A Ras-related protein Rab-8A KLEGNSPQGSNQGVK S181/S185 
B3KVN0 SLC2A1 Solute carrier family 2, facilitated glucose transporter member 1 TPEELFHPLGADSQV S490 
A0A087WYB4 STOML2 Stomatin-like protein 2, mitochondrial APVPGTPDSLSSGSSR T327 
Q8IV54 TSC22D4 TSC22D4 protein NGSPPPGAPSSR S62 
P16157 ANK1 Ankyrin-1 NGASPNEVSSDGTTPLAIAK S759 
A0A0U1RQT1 ACAP2 Arf-GAP with coiled-coil, ANK repeat, and PH domain-containing protein 2 YSISLSPPEQQK S521 
P27824 CANX Calnexin QKSDAEEDGGTVSQEEEDR S564 
G3V238 EEF1AKMT2 EEF1A lysine methyltransferase 2 SDKGSPGEDGFVPSALGTR S21 
P05198 EIF2S1 Eukaryotic translation initiation factor 2 subunit 1 VVTDTDETELAR T279 
F8WAE5 EIF2A Eukaryotic translation initiation factor 2A SDKSPDLAPTPAPQSTPR S506 
C9JII4 GCFC2 GC-rich sequence DNA-binding factor 2 ELPVPGSAEEEPPSGGGR S40 
Q2VPJ6 HSP90AA1 HSP90AA1 protein DKEVSDDEAEEK S231 
D3DWY2 IKBKG Inhibitor of κ light polypeptide gene enhancer in B-cells, kinase γ SPPEEPPDFCCPK S387 
O60307 MAST3 Microtubule-associated serine/threonine-protein kinase 3 SSENVLDEEGGR S134 
F2Z2K0 NSFL1C NSFL1 cofactor p47 SPNELVDDLFK S114 
B7Z8Y1 NFATC2 Nuclear factor of activated T-cells, cytoplasmic 2 TSPDPSPVSAAPSK S330 
H0Y5Q9 INPP5D Phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase 1 GESPPTPPGQPPISPK T963 
D6RC77 PGM3 Phosphoacetylglucosamine mutase STIGVMVTASHNPEEDNGVK S64 
M4QHP2 PIEZO1 Piezo-type mechanosensitive ion channel component SGSEEAVTDPGER S1621 
Q9BVG4 PBDC1 Protein PBDC1 GADSGEEKEEGINR S197 
B4E3V2 SQSTM1 Sequestosome-1 LTPVSPESSSTEEK T269/S272 
Q6PJD5 SHARPIN SHARPIN protein SPGNLTER S165 
B2RMN7 SPTB Spectrin β chain TSPVSLWSR S2114/S2117 
C9JWF0 SMC4 Structural maintenance of chromosomes protein 4 REEGPPPPSPDGASSDAEPEPPSGR S22 
H0YN01 TLN2 Talin-2 LDEGTPPEPK T1843 
C9J3F6 TBC1D5 TBC1 domain family member 5 NISSSPSVESLPGGR S541/S544 
E9PJJ0 TBCEL Tubulin-specific chaperone cofactor E-like protein YSPENFPYR S18 
Q8IXQ3 C9orf40 Uncharacterized protein C9orf40 RDSGDNSAPSGQER S76 
Q5JSH3 WDR44 WD repeat-containing protein 44 VGNESPVQELK S50 
F8WBS8 PSMD2 26S proteasome non-ATPase regulatory subunit 2 DKAPVQPQQSPAAAPGGTDEKPSGK S16 
Q92625 ANKS1A Ankyrin repeat and SAM domain-containing protein 1A SESLSNCSIGK S647 
E9PJP4 EPB41L2 Band 4.1-like protein 2 EVAENQQNQSSDPEEEKGSQPPPAAESQSSLR S39/S58 
A8MZ77 BSG Basigin RKPEDVLDDDDAGSAPLK S362 
Q9NSY1 BMP2K BMP-2-inducible protein kinase DSQSSNEFLTISDSK S1029 
A0A0C4DGD1 CAST Calpastatin DTSQSDKDLDDALDK S563 
Q9Y5K6 CD2AP CD2-associated protein KNSLDELR S582 
Q8TEH3 DENND1A DENN domain-containing protein 1A TGGTLSDPEVQR T24 
B4DU91 EPN1 Epsin-1 SPGAFDMSGVR S435 
M0QXW7 KHSRP Far upstream element-binding protein 2 VQISPDSGGLPER S181 
F5H1C6 FERMT3 Fermitin family homolog 3 TASGDYIDSSWELR Y11 
P04075 ALDOA Fructose-bisphosphate aldolase A GILAADESTGSIAKR S36 
H0YJ30 GPHN Gephyrin ASHSAVDITK S303/S305 
Q2Q9B7 G6PD Glucose-6-phosphate 1-dehydrogenase VQPNEAVYTK Y401 
E9PK89 RAB3IL1 Guanine nucleotide exchange factor for Rab-3A ELHPQLLSPTK S179 
O75633 GNAS Guanine nucleotide-binding protein Gs α subunit isoform L3 ISTASGDGR S352 
Q5H924 HUWE1 HECT, UBA, and WWE domain containing 1 REESPMDVDQPSPSAQDTQSIASDGTPQGEK S3816 
A0A0F7NGI8 LRRFIP1 Leucine rich repeat IDGATQSSPAEPK S714 
E9PJF4 CLNS1A Methylosome subunit pICln EPVADEEEEDSDDDVEPITEFR S102 
S4R3D6 APIP Methylthioribulose-1-phosphate dehydratase DISGPSPSKK S87 
Q6NT16 SLC18B1 MFS-type transporter SLC18B1 SKSQNILSTEEER S438 
B4DS42 OATL1 Ornithine aminotransferase-like 1 QASLDGLQQLR S506 
P30041 PRDX6 Peroxiredoxin-6 DFTPVCTTELGR T44 
I3L1U0 RILP Rab-interacting lysosomal protein AESSEDETSSPAPSK S354 
Q9P227 ARHGAP23 ρ GTPase-activating protein 23 SAEALGPGALVSPR S361/S372 
Q16513 PKN2 Serine/threonine-protein kinase N2 ASSLGEIDESSELR S583 
C9JIG9 OXSR1 Serine/threonine-protein kinase OSR1 TEDGGWEWSDDEFDEESEEGK S339 
P02549 SPTA1 Spectrin α chain, erythrocytic 1 QDTLDASLQSFQQER S1976 
B2RMN7 SPTB Spectrin β chain LSSSWESLQPEPSHPY S2123/S2125/S2128 
B4DZC9 SLK STE20-like serine/threonine-protein kinase VDEDSAEDTQSNDGK S571 
P17987 TCP1 T-complex protein 1 subunit α HGSYEDAVHSGALND S544/Y545 
B4DUI5 TPI1 Triosephosphate isomerase IIYGGSVTGATCK Y246 
Q8IV54 TSC22D4 TSC22D4 protein VEAEAGGSGARTPPLSR T229 
B4DQ79 LYN Tyrosine-protein kinase VIEDNEYTAR Y397 
A0A024R8I2 UBADC1 Ubiquitin associated domain containing 1, isoform CRA_c APSPLPK S98 
Q5VVQ6 YOD1 Ubiquitin thioesterase OTU1 SSPAFTK S130 
B3KW31 VPS13A Vacuolar protein sorting 13A QASFTDVRDPSLK S1416 
A0A0S2Z3K9 CRK V-crk sarcoma virus CT10 oncogene-like protein isoform 2 DSSTSPGDYVLSVSENSR S41 
Accession no.Gene symbolProtein nameSequenceCountSite
E9PIJ1 AMPD2 AMP deaminase 2 TDSDSDLQLYK 7* S190/S197 
A0A0U1RQT1 ACAP2 Arf-GAP with coiled-coil, ANK repeat, and PH domain-containing protein 2 SSPSTGSLDSGNESK 7* S384 
B7Z3I9 ALAD Δ-aminolevulinic acid dehydratase SSPAFGDR S215 
P49840 GSK3A Glycogen synthase kinase-3 α GEPNVSYICSR 7* Y279 
P49840 GSK3A Glycogen synthase kinase-3 α TSSFAEPGGGGGGGGGGPGGSASGPGGTGGGK 7* S21/S39 
E9PK89 RAB3IL1 Guanine nucleotide exchange factor for Rab-3A TLVITSTPASPNR S168 
B4DMA2 HSP90AB1 Heat shock protein HSP 90-β IEDVGSDEEDDSGKDK 7* S255 
Q9H1E3 NUCKS1 Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 VVDYSQFQESDDADEDYGR 7* S19 
S4R2Y4 CBX3 Chromobox protein homolog 3 SLSDSESDDSK S93 
Q08495 DMTN Dematin DSSVPGSPSSIVAK S26 
Q9UG54 MAP3K7 Mitogen-activated protein kinase kinase kinase 7 SIQDLTVTGTEPGQVSSR S439 
P61006 RAB8A Ras-related protein Rab-8A KLEGNSPQGSNQGVK S181/S185 
B3KVN0 SLC2A1 Solute carrier family 2, facilitated glucose transporter member 1 TPEELFHPLGADSQV S490 
A0A087WYB4 STOML2 Stomatin-like protein 2, mitochondrial APVPGTPDSLSSGSSR T327 
Q8IV54 TSC22D4 TSC22D4 protein NGSPPPGAPSSR S62 
P16157 ANK1 Ankyrin-1 NGASPNEVSSDGTTPLAIAK S759 
A0A0U1RQT1 ACAP2 Arf-GAP with coiled-coil, ANK repeat, and PH domain-containing protein 2 YSISLSPPEQQK S521 
P27824 CANX Calnexin QKSDAEEDGGTVSQEEEDR S564 
G3V238 EEF1AKMT2 EEF1A lysine methyltransferase 2 SDKGSPGEDGFVPSALGTR S21 
P05198 EIF2S1 Eukaryotic translation initiation factor 2 subunit 1 VVTDTDETELAR T279 
F8WAE5 EIF2A Eukaryotic translation initiation factor 2A SDKSPDLAPTPAPQSTPR S506 
C9JII4 GCFC2 GC-rich sequence DNA-binding factor 2 ELPVPGSAEEEPPSGGGR S40 
Q2VPJ6 HSP90AA1 HSP90AA1 protein DKEVSDDEAEEK S231 
D3DWY2 IKBKG Inhibitor of κ light polypeptide gene enhancer in B-cells, kinase γ SPPEEPPDFCCPK S387 
O60307 MAST3 Microtubule-associated serine/threonine-protein kinase 3 SSENVLDEEGGR S134 
F2Z2K0 NSFL1C NSFL1 cofactor p47 SPNELVDDLFK S114 
B7Z8Y1 NFATC2 Nuclear factor of activated T-cells, cytoplasmic 2 TSPDPSPVSAAPSK S330 
H0Y5Q9 INPP5D Phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase 1 GESPPTPPGQPPISPK T963 
D6RC77 PGM3 Phosphoacetylglucosamine mutase STIGVMVTASHNPEEDNGVK S64 
M4QHP2 PIEZO1 Piezo-type mechanosensitive ion channel component SGSEEAVTDPGER S1621 
Q9BVG4 PBDC1 Protein PBDC1 GADSGEEKEEGINR S197 
B4E3V2 SQSTM1 Sequestosome-1 LTPVSPESSSTEEK T269/S272 
Q6PJD5 SHARPIN SHARPIN protein SPGNLTER S165 
B2RMN7 SPTB Spectrin β chain TSPVSLWSR S2114/S2117 
C9JWF0 SMC4 Structural maintenance of chromosomes protein 4 REEGPPPPSPDGASSDAEPEPPSGR S22 
H0YN01 TLN2 Talin-2 LDEGTPPEPK T1843 
C9J3F6 TBC1D5 TBC1 domain family member 5 NISSSPSVESLPGGR S541/S544 
E9PJJ0 TBCEL Tubulin-specific chaperone cofactor E-like protein YSPENFPYR S18 
Q8IXQ3 C9orf40 Uncharacterized protein C9orf40 RDSGDNSAPSGQER S76 
Q5JSH3 WDR44 WD repeat-containing protein 44 VGNESPVQELK S50 
F8WBS8 PSMD2 26S proteasome non-ATPase regulatory subunit 2 DKAPVQPQQSPAAAPGGTDEKPSGK S16 
Q92625 ANKS1A Ankyrin repeat and SAM domain-containing protein 1A SESLSNCSIGK S647 
E9PJP4 EPB41L2 Band 4.1-like protein 2 EVAENQQNQSSDPEEEKGSQPPPAAESQSSLR S39/S58 
A8MZ77 BSG Basigin RKPEDVLDDDDAGSAPLK S362 
Q9NSY1 BMP2K BMP-2-inducible protein kinase DSQSSNEFLTISDSK S1029 
A0A0C4DGD1 CAST Calpastatin DTSQSDKDLDDALDK S563 
Q9Y5K6 CD2AP CD2-associated protein KNSLDELR S582 
Q8TEH3 DENND1A DENN domain-containing protein 1A TGGTLSDPEVQR T24 
B4DU91 EPN1 Epsin-1 SPGAFDMSGVR S435 
M0QXW7 KHSRP Far upstream element-binding protein 2 VQISPDSGGLPER S181 
F5H1C6 FERMT3 Fermitin family homolog 3 TASGDYIDSSWELR Y11 
P04075 ALDOA Fructose-bisphosphate aldolase A GILAADESTGSIAKR S36 
H0YJ30 GPHN Gephyrin ASHSAVDITK S303/S305 
Q2Q9B7 G6PD Glucose-6-phosphate 1-dehydrogenase VQPNEAVYTK Y401 
E9PK89 RAB3IL1 Guanine nucleotide exchange factor for Rab-3A ELHPQLLSPTK S179 
O75633 GNAS Guanine nucleotide-binding protein Gs α subunit isoform L3 ISTASGDGR S352 
Q5H924 HUWE1 HECT, UBA, and WWE domain containing 1 REESPMDVDQPSPSAQDTQSIASDGTPQGEK S3816 
A0A0F7NGI8 LRRFIP1 Leucine rich repeat IDGATQSSPAEPK S714 
E9PJF4 CLNS1A Methylosome subunit pICln EPVADEEEEDSDDDVEPITEFR S102 
S4R3D6 APIP Methylthioribulose-1-phosphate dehydratase DISGPSPSKK S87 
Q6NT16 SLC18B1 MFS-type transporter SLC18B1 SKSQNILSTEEER S438 
B4DS42 OATL1 Ornithine aminotransferase-like 1 QASLDGLQQLR S506 
P30041 PRDX6 Peroxiredoxin-6 DFTPVCTTELGR T44 
I3L1U0 RILP Rab-interacting lysosomal protein AESSEDETSSPAPSK S354 
Q9P227 ARHGAP23 ρ GTPase-activating protein 23 SAEALGPGALVSPR S361/S372 
Q16513 PKN2 Serine/threonine-protein kinase N2 ASSLGEIDESSELR S583 
C9JIG9 OXSR1 Serine/threonine-protein kinase OSR1 TEDGGWEWSDDEFDEESEEGK S339 
P02549 SPTA1 Spectrin α chain, erythrocytic 1 QDTLDASLQSFQQER S1976 
B2RMN7 SPTB Spectrin β chain LSSSWESLQPEPSHPY S2123/S2125/S2128 
B4DZC9 SLK STE20-like serine/threonine-protein kinase VDEDSAEDTQSNDGK S571 
P17987 TCP1 T-complex protein 1 subunit α HGSYEDAVHSGALND S544/Y545 
B4DUI5 TPI1 Triosephosphate isomerase IIYGGSVTGATCK Y246 
Q8IV54 TSC22D4 TSC22D4 protein VEAEAGGSGARTPPLSR T229 
B4DQ79 LYN Tyrosine-protein kinase VIEDNEYTAR Y397 
A0A024R8I2 UBADC1 Ubiquitin associated domain containing 1, isoform CRA_c APSPLPK S98 
Q5VVQ6 YOD1 Ubiquitin thioesterase OTU1 SSPAFTK S130 
B3KW31 VPS13A Vacuolar protein sorting 13A QASFTDVRDPSLK S1416 
A0A0S2Z3K9 CRK V-crk sarcoma virus CT10 oncogene-like protein isoform 2 DSSTSPGDYVLSVSENSR S41 

List of significantly enriched or exclusive phosphopeptides in reticulocytes after deformation, as assayed through microsphiltration and microcapillary traversal. Generally, only phosphopeptides present in at least 4 of the 7 postdeformation samples (4 microsphiltration + 3 microfluidics) and not present in at least 4 of the 7 predeformation samples were included. The exception to this analysis consists of the phosphopeptides counted as 7*, which were present in all postdeformation samples but also in 4 of 7 of the predeformation samples. The prediction of the most probable phosphosites in the phosphopeptide was included according to the count of occurrences in previous low-throughput and high-throughput studies, as assayed through PhosphoSitePlus.50 

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