Table 1.
List of ADP-ribosylated peptides identified from LPS/U0126 stimulated microglia.
| gene | protein | protein accession | ADPR site | ADPR AA | confidence | peptide sequence | peptide score | expect value |
|---|---|---|---|---|---|---|---|---|
| Alcam | CD166 antigen | tr|E9Q3Q6|E9Q3Q6_MOUSE | 319 | R | 100% | (R)NMAASTTITVHYLDLSLNPSGEVTK | 54 | 0,0015 |
| Alcam | CD166 antigen | tr|E9Q3Q6|E9Q3Q6_MOUSE | 386 | D | 99,85% | LRSSPSFSSLHYQ(D)AGNYVCETALQEVEGLK | 74,34 | 2,20E-07 |
| Bcam | Basal cell adhesion molecule | sp|Q9R069|BCAM_MOUSE | 484 | D | 96,83% | LTWSQRG(D)TPAEPPFEGR | 30,05 | 0,0031 |
| Cd14 | Monocyte differentiation antigen CD14 | sp|P10810|CD14_MOUSE | 68 | R | 100% | (R)VDTEADLGQFTDIIK | 132,14 | 2,00E-11 |
| Cd40 | Isoform V of Tumor necrosis factor receptor superfamily member 5 | sp|P27512-5|TNR5_MOUSE | 247 | E | 99,83% | ISVQ(E)RQVTDSIALRPLV | 52,58 | 1,20E-05 |
| Cd9 | CD9 antigen | sp|P40240|CD9_MOUSE | 131 | R | 96,99% | L(R)SKDEPQR | 27,61 | 0,0053 |
| Clec12a | C-type lectin domain family 12 member A | sp|Q504P2|CL12A_MOUSE | 197 | D | 100% | K(D)RTQYPLSEK | 32,74 | 0,0018 |
| Clec12a | C-type lectin domain family 12 member A | sp|Q504P2|CL12A_MOUSE | 215 | R | 97,79% | MFLSEESE(R)STDDIDKK | 27,02 | 0,0064 |
| Enpp1 | Ectonucleotide pyrophosphatase/phosphodiesterase 1 isoform CRA_d | tr|G3 × 9S2|G3 × 9S2_MOUSE | 853 | R | 100% | (R)ESSWVEELLTLHR | 96,1 | 7,40E-08 |
| Enpp1 | Ectonucleotide pyrophosphatase/phosphodiesterase 1 isoform CRA_d | tr|G3 × 9S2|G3 × 9S2_MOUSE | 854 | E | 99,31% | R(E)SSWVEELLTLHR | 63,12 | 1,50E-06 |
| Fcer1g | High affinity immunoglobulin epsilon receptor subunit gamma | sp|P20491|FCERG_MOUSE | 71 | R | 100,00% | EKADAVYTGLNT(R)SQETYETLK | 76,36 | 7,90E-06 |
| Fcgr1 | High affinity immunoglobulin gamma Fc receptor I | sp|P26151|FCGR1_MOUSE | 184 | R | 100% | H(R)YTSAGVSITVK | 40,29 | 0,00025 |
| Fcgr2b | Low affinity immunoglobulin gamma Fc region receptor II | tr|E9Q415|E9Q415_MOUSE | 198 | R | 100,00% | GSLG(R)TLHQSKPVTITVQGPK | 107,45 | 9,70E-11 |
| Fcgr2b | Low affinity immunoglobulin gamma Fc region receptor II | tr|E9Q415|E9Q415_MOUSE | 88 | R | 100% | SI(R)SQVQASYTFK | 61,23 | 1,80E-06 |
| Fcgr2b | Low affinity immunoglobulin gamma Fc region receptor II | tr|E9Q415|E9Q415_MOUSE | 158 | R | 100% | LLN(R)ISFFHNEK | 30,35 | 0,0016 |
| H2-D1 | Isoform 2 of H-2 class I histocompatibility antigen alpha chain | sp|P01895-2|HA1Y_MOUSE | 7 | R | 100% | A(R)WIEQEGPEYWER | 77,61 | 4,10E-06 |
| H2-K1 | H-2 class I histocompatibility antigen K-D alpha chain | sp|P01902|HA1D_MOUSE | 169 | E | 98,50% | RKW(E)QAGDAEYYR | 31,46 | 0,0011 |
| Hfe | Hemochromatosis isoform CRA_a | tr|Q5SZ87|Q5SZ87_MOUSE | 120 | E | 100% | LL(E)LGRGVLGQQVPTLVK | 47,31 | 7,20E-05 |
| Icam1 | Isoform 2 of Intercellular adhesion molecule 1 | sp|P13597-2|ICAM1_MOUSE | 345 | R | 99,86% | VVLLSGVEP(R)PPTPQVQFTLNASSEDHKR | 35,05 | 0,00054 |
| Icam1 | Isoform 2 of Intercellular adhesion molecule 1 | sp|P13597-2|ICAM1_MOUSE | 152 | R | 99,02% | GEEILS(R)QPVGGHPK | 27,87 | 0,0057 |
| Icoslg | Isoform 2 of ICOS ligand | sp|Q9JHJ8-2|ICOSL_MOUSE | 108 | R | 100% | N(R)GHLSLDSMK | 30,38 | 0,0029 |
| Il2rg | Cytokine receptor common subunit gamma | sp|P34902|IL2RG_MOUSE | 196 | R | 100% | D(R)SWTELIVNHEPR | 41,46 | 0,00013 |
| Itga5 | Integrin alpha-5 | sp|P11688|ITA5_MOUSE | 903 | R | 100% | EAPG(R)SSTASGTQVLK | 25,81 | 0,0064 |
| Itga6 | Integrin alpha-6 (Fragment) | tr|F6VSK8|F6VSK8_MOUSE | 188 | R | 100% | LRPIPITASVEIQEPSS(R) | 69,51 | 2,70E-05 |
| Itgam | Integrin alpha-M | tr|Q3U1U4|Q3U1U4_MOUSE | 415 | R | 100% | N(R)VQSLVLGAPR | 51,11 | 2,00E-05 |
| Itgam | Integrin alpha-M | tr|Q3U1U4|Q3U1U4_MOUSE | 502 | R | 100% | A(R)WQCEALLHGDQGHPWGR | 94,4 | 1,20E-07 |
| Itgam | Integrin alpha-M | tr|Q3U1U4|Q3U1U4_MOUSE | 277 | R | 99,18% | FGDPLDYKDVIPEAD(R)AGVIR | 50,86 | 4,20E-05 |
| Itgam | Integrin alpha-M | tr|Q3U1U4|Q3U1U4_MOUSE | 506 | E | 95,00% | ARWQC(E)ALLHGDQGHPWGR | 30,38 | 0,0017 |
| Itgav | Integrin alpha-V light chain | tr|A2AKI5|A2AKI5_MOUSE | 855 | R | 100% | DLTL(R)EGDVHTLGCGIAK | 60,04 | 0,00031 |
| Itgb1 | Isoform 2 of Integrin beta-1 | sp|P09055-2|ITB1_MOUSE | 228 | R | 100% | NVLSLTD(R)GEFFNELVGQQR | 55,9 | 7,50E-06 |
| Itgb2 | Integrin beta | tr|Q542I8|Q542I8_MOUSE | 454 | R | 100% | DQS(R)EQSLCGGK | 41,49 | 0,0002 |
| Itgb5 | Integrin beta | tr|Q6PE70|Q6PE70_MOUSE | 226 | R | 100% | HLLPLTD(R)VDSFNEEVRK | 30,98 | 0,0013 |
| Itgb5 | Integrin beta | tr|Q6PE70|Q6PE70_MOUSE | 72 | R | 100% | ANLI(R)NGCEGEIESPASSTHVLR | 84,68 | 1,30E-06 |
| Itgb5 | Integrin beta | tr|Q6PE70|Q6PE70_MOUSE | 158 | R | 99,85% | DDLENI(R)SLGTK | 39,92 | 0,031 |
| Lgals1 | Galectin-1 | sp|P16045|LEG1_MOUSE | 116 | E | 99,97% | FPNRLNM(E)AINYMAADGDFK | 42,7 | 0,00015 |
| Lrpap1 | Alpha-2-macroglobulin receptor-associated protein | sp|P55302|AMRP_MOUSE | 71 | R | 100% | (R)LHLSPVR | 20,46 | 0,012 |
| Mcam | Isoform 2 of Cell surface glycoprotein MUC18 | sp|Q8R2Y2-2|MUC18_MOUSE | 71 | R | 100% | E(R)QILIFR | 39,14 | 0,0066 |
| Mrc1 | Macrophage mannose receptor 1 | sp|Q61830|MRC1_MOUSE | 545 | R | 100,00% | AYLTTVED(R)YEQAFLTSLVGLRPEK | 85,03 | 1,10E-08 |
| Pdia3 | Protein disulfide-isomerase A3 | sp|P27773|PDIA3_MOUSE | 62 | R | 100% | (R)LAPEYEAAATR | 63,08 | 0,00013 |
| Ptprc | Receptor-type tyrosine-protein phosphatase C | tr|S4R1M0|S4R1M0_MOUSE | 168 | R | 100% | (R)NTFIPER | 24,41 | 0,0051 |
| Ptprc | Receptor-type tyrosine-protein phosphatase C | tr|S4R1M0|S4R1M0_MOUSE | 508 | R | 100% | N(R)YVDILPYDYNR | 42 | 0,015 |
| Ptprj | Receptor-type tyrosine-protein phosphatase eta | sp|Q64455|PTPRJ_MOUSE | 202 | R | 100% | (R)IPVTNLSQLHK | 29,43 | 0,0017 |
| Pvrl2 | Isoform Alpha of Nectin-2 | sp|P32507-2|PVRL2_MOUSE | 104 | R | 100% | A(R)PETNADLR | 20 | 0,037 |
| Sdc3 | Syndecan-3 | sp|Q64519|SDC3_MOUSE | 362 | D | 98,15% | GARPGPGLH(D)NAIDSGSSAAQLPQK | 45,2 | 0,00037 |
| Sdc4 | Syndecan-4 | sp|O35988|SDC4_MOUSE | 116 | R | 100% | (R)APSDVGDDMSNK | 35,64 | 0,023 |
| Slamf7 | Isoform 4 of SLAM family member 7 | sp|Q8BHK6-4|SLAF7_MOUSE | 77 | R | 100% | E(R)IVFPDGLYSMK | 20,04 | 0,013 |
| Slc3a2 | Isoform 2 of 4F2 cell-surface antigen heavy chain | sp|P10852-2|4F2_MOUSE | 509 | R | 100% | SA(R)LGASNLPAGISLPASAK | 52,26 | 0,00024 |
| Slc3a2 | Isoform 2 of 4F2 cell-surface antigen heavy chain | sp|P10852-2|4F2_MOUSE | 168 | R | 99,99% | IGDLQAFVG(R)DAGGIAGLK | 46,05 | 0,0073 |
| Slc9a1 | Sodium/hydrogen exchanger 1 | sp|Q61165|SL9A1_MOUSE | 622 | R | 100% | AVTSD(R)ILPALSK | 33,64 | 0,005 |
| Stx4 | Syntaxin-4 | sp|P70452|STX4_MOUSE | 192 | D | 99,99% | (D)TQVTRQALNEISAR | 52,29 | 2,90E-05 |
| Tfrc | Transferrin receptor protein 1 | sp|Q62351|TFR1_MOUSE | 658 | R | 100% | ATS(R)LTTDFHNAEK | 31,69 | 0,007 |
| Tfrc | Transferrin receptor protein 1 | sp|Q62351|TFR1_MOUSE | 358 | R | 100% | MEGSCPA(R)WNIDSSCK | 41,06 | 0,029 |
| Tlr2 | Toll-like receptor | tr|G3 × 8Y8|G3 × 8Y8_MOUSE | 187 | R | 100% | ALSL(R)NYQSQSLK | 32,78 | 0,0011 |
| Vamp3 | Vesicle-associated membrane protein 3 | sp|P63024|VAMP3_MOUSE | 18 | R | 100% | (R)LQQTQNQVDEVVDIMR | 57,88 | 0,00054 |
Mixed glial cells were stimulated with LPS (0.1 µg/ml) and U0126 (10 µM) overnight in BME medium containing 10% FCS. CD11b+ microglia were FACS sorted, incubated with 50 µM NAD and 2.5 mM DTT for 30 min on ice, washed twice with PBS and lysed using RIPA buffer. After trypsin digestion ADPR peptides were enriched using macrodomain Af1521 and samples were subjected to mass spectrometry analyses. Only peptides of cell surface proteins displaying a peptide score >20 and a confidence of >95% were included into the analyses.