UniProt UniProt http://www.uniprot.org Induction of myelodysplasia by myeloid-derived suppressor cells. Induction of myelodysplasia by myeloid-derived suppressor cells. The Journal of clinical investigation 2013 imex curation Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org Accepted 2019-MAY-16 AT 13:09 BST AT 13:09 BST by MGT The Journal of clinical investigation 2013 Only protein-protein interactions imex curation imex curation In the absence of any other information, I was going with coip because of the use of the word immunopreciptiation in the text, but have gone up a couple of nodes to affinity chromatography since this is open to doubt.; I think CD33 is probably full length so have gone with that, Not clear where S100A9 is expressed. Induction of myelodysplasia by myeloid-derived suppressor cells. Induction of myelodysplasia by myeloid-derived suppressor cells. The Journal of clinical investigation 2013 imex curation Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom western blot western blot Immuno blot Immuno blot Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org Accepted 2019-MAY-16 AT 13:09 BST AT 13:09 BST by MGT The Journal of clinical investigation 2013 Only protein-protein interactions imex curation imex curation In the absence of any other information, I was going with coip because of the use of the word immunopreciptiation in the text, but have gone up a couple of nodes to affinity chromatography since this is open to doubt.; I think CD33 is probably full length so have gone with that, Not clear where S100A9 is expressed. Induction of myelodysplasia by myeloid-derived suppressor cells. Induction of myelodysplasia by myeloid-derived suppressor cells. The Journal of clinical investigation 2013 imex curation Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-bone marrow Human bone marrow bone_marrow bone marrow anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org Accepted 2019-MAY-16 AT 13:09 BST AT 13:09 BST by MGT The Journal of clinical investigation 2013 Only protein-protein interactions imex curation imex curation In the absence of any other information, I was going with coip because of the use of the word immunopreciptiation in the text, but have gone up a couple of nodes to affinity chromatography since this is open to doubt.; I think CD33 is probably full length so have gone with that, Not clear where S100A9 is expressed. Induction of myelodysplasia by myeloid-derived suppressor cells. Induction of myelodysplasia by myeloid-derived suppressor cells. The Journal of clinical investigation 2013 imex curation Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-sjcrh30 Human bone marrow rhabdomyosarcoma SJCH30 cell sjcrh30 Bone marrow rhabdomyosarcoma SJCRH30 cells elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org Accepted 2019-MAY-16 AT 13:09 BST AT 13:09 BST by MGT The Journal of clinical investigation 2013 Only protein-protein interactions imex curation imex curation In the absence of any other information, I was going with coip because of the use of the word immunopreciptiation in the text, but have gone up a couple of nodes to affinity chromatography since this is open to doubt.; I think CD33 is probably full length so have gone with that, Not clear where S100A9 is expressed. Induction of myelodysplasia by myeloid-derived suppressor cells. Induction of myelodysplasia by myeloid-derived suppressor cells. The Journal of clinical investigation 2013 imex curation Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-sjcrh30 Human bone marrow rhabdomyosarcoma SJCH30 cell sjcrh30 Bone marrow rhabdomyosarcoma SJCRH30 cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org Accepted 2019-MAY-16 AT 13:09 BST AT 13:09 BST by MGT The Journal of clinical investigation 2013 Only protein-protein interactions imex curation imex curation In the absence of any other information, I was going with coip because of the use of the word immunopreciptiation in the text, but have gone up a couple of nodes to affinity chromatography since this is open to doubt.; I think CD33 is probably full length so have gone with that, Not clear where S100A9 is expressed. Induction of myelodysplasia by myeloid-derived suppressor cells. Induction of myelodysplasia by myeloid-derived suppressor cells. The Journal of clinical investigation 2013 imex curation Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. sheng.wei@moffitt.org 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom sequence tag sequence tag identification sequence tag Chen X., Eksioglu EA., Zhou J., Zhang L., Djeu J., Fortenbery N., Epling-Burnette P., Van Bijnen S., Dolstra H., Cannon J., Youn JI., Donatelli SS., Qin D., De Witte T., Tao J., Wang H., Cheng P., Gabrilovich DI., List A., Wei S. Bone Marrow cell lysates from patients with MDS (Myelodysplastic syndromes) sheng.wei@moffitt.org Accepted 2019-MAY-16 AT 13:09 BST AT 13:09 BST by MGT The Journal of clinical investigation 2013 Only protein-protein interactions imex curation imex curation In the absence of any other information, I was going with coip because of the use of the word immunopreciptiation in the text, but have gone up a couple of nodes to affinity chromatography since this is open to doubt.; I think CD33 is probably full length so have gone with that, Not clear where S100A9 is expressed. cd33_human Myeloid cell surface antigen CD33 CD33 SIGLEC3 Sialic acid-binding Ig-like lectin 3 gp67 protein protein human Homo sapiens Human MPLLLLLPLLWAGALAMDPNFWLQVQESVTVQEGLCVLVPCTFFHPIPYYDKNSPVHGYWFREGAIISRDSPVATNKLDQEVQEETQGRFRLLGDPSRNNCSLSIVDARRRDNGSYFFRMERGSTKYSYKSPQLSVHVTDLTHRPKILIPGTLEPGHSKNLTCSVSWACEQGTPPIFSWLSAAPTSLGPRTTHSSVLIITPRPQDHGTNLTCQVKFAGAGVTTERTIQLNVTYVPQNPTTGIFPGDGSGKQETRAGVVHGAIGGAGVTALLALCLCLIFFIVKTHRRKAARTAVGRNDTHPTTGSASPKHQKKSKLHGPTETSSCSGAAPTVEMDEELHYASLNFHGMNPSKDTSTEYSEVRTQ 1C73E588240FBAD8 s10a9_human Protein S100-A9 S100 calcium-binding protein A9 Calgranulin-B Migration inhibitory factor-related protein 14 Leukocyte L1 complex heavy chain Calprotectin L1H subunit S100A9 CAGB CFAG MRP14 protein protein human Homo sapiens Human MTCKMSQLERNIETIINTFHQYSVKLGHPDTLNQGEFKELVRKDLQNFLKKENKNEKVIEHIMEDLDTNADKQLSFEEFIMLMARLTWASHEKMHEGDEGPGHHHKPGLGEGTP C3BE19729E14C078 ptn6_human Tyrosine-protein phosphatase non-receptor type 6 PTPN6 HCP Protein-tyrosine phosphatase 1C Hematopoietic cell protein-tyrosine phosphatase SH-PTP1 Protein-tyrosine phosphatase SHP-1 PTP1C protein protein human Homo sapiens Human MVRWFHRDLSGLDAETLLKGRGVHGSFLARPSRKNQGDFSLSVRVGDQVTHIRIQNSGDFYDLYGGEKFATLTELVEYYTQQQGVLQDRDGTIIHLKYPLNCSDPTSERWYHGHMSGGQAETLLQAKGEPWTFLVRESLSQPGDFVLSVLSDQPKAGPGSPLRVTHIKVMCEGGRYTVGGLETFDSLTDLVEHFKKTGIEEASGAFVYLRQPYYATRVNAADIENRVLELNKKQESEDTAKAGFWEEFESLQKQEVKNLHQRLEGQRPENKGKNRYKNILPFDHSRVILQGRDSNIPGSDYINANYIKNQLLGPDENAKTYIASQGCLEATVNDFWQMAWQENSRVIVMTTREVEKGRNKCVPYWPEVGMQRAYGPYSVTNCGEHDTTEYKLRTLQVSPLDNGDLIREIWHYQYLSWPDHGVPSEPGGVLSFLDQINQRQESLPHAGPIIVHCSAGIGRTGTIIVIDMLMENISTKGLDCDIDIQKTIQMVRAQRSGMVQTEAQYKFIYVAIAQFIETTKKKLEVLQSQKGQESEYGNITYPPAMKNAHAKASRTSSKHKEDVYENLHTKNKREEKVKKQRSADKEKSKGSLKRK DIP protein P29350 original sequence version: 183 DIP protein P29350 original sequence version: 139 DIP protein P29350-1 original sequence version: 139 4D7736C21D3542D2 cd33-s100a9-1 6 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-bone marrow Human bone marrow bone_marrow bone marrow 7 unspecified role unspecified role bait bait purified purified ectodomain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. physical association physical association Fig. 3A BM cell lysates from MDS patients were immunoprecipitated with the CD33-fusion followed by high-throughput mass-spectrometric analysis on associated peptides. S100A9, an inflammatory signaling molecule known to activate MDSC, was among the most prominent protein bands identified (Figure ​(Figure3A).3A). s100a9-cd33-1 4 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed human-sjcrh30 Human bone marrow rhabdomyosarcoma SJCH30 cell sjcrh30 Bone marrow rhabdomyosarcoma SJCRH30 cells 7 unspecified role unspecified role prey prey ectodomain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. proximity proximity close-contact colocalization neighbourhood interaction Fig. 3C Direct binding of S100A9 to CD33 was confirmed further in a sandwich ELISA where the capture antibody was anti-S100A9 (Figure ​(Figure3C)3C) s100a9-cd33-2 5 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association Fig. 3E cd33-s100a9-2 2 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed human-bone marrow Human bone marrow bone_marrow bone marrow 7 unspecified role unspecified role bait bait purified purified ectodomain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. physical association physical association Fig. 3F The highest amount of S100A9 was precipitated from the BM of Myelodysplastic syndromes patient specimens To fully corroborate the binding of this pair, we set out to perform a reverse immunoprecipitation on CD33 transfected cells showing that S100A9 coprecipitated with CD33 only in recombinant human S100A9–treated (rhS100A9-treated) cells (Figure ​(Figure3E).3E). cd33-ptpn6-5 3 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig. 3N S100A9 (P06702) Bone marrow isolated from Myelodysplastic syndromes patients rhS100A9 ligation correspondingly increased the recruitment of SHP-1, confirming that CD33 signals through its ITIM after S100A9 ligation in MDS BM (Figure ​(Figure3N).3N) cd33-ptpn6-6 5 9 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Fig. 3O cd33-s100a9-3 1 8 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 tag fluorescence tag visualisation by fluorescence tag fluorescence immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed human-mdsc Human myeloid-derived suppressor cells mdsc Myeloid-derived suppressor cell colocalization colocalization Fig. 3M BM extracted from Myelodysplastic syndromes patient specimens Moreover, the engagement of CD33 with rhS100A9 in MDS-MDSC from patient BM resulted in a time-dependent colocalization of this ligand/receptor pair (Figure ​(Figure3M).3M).