Ser221

Site Information
kVAAEtQsPsLFGst SwissProt Entrez-Gene
Blast this site against: NCBI SwissProt PDB
Site Group ID: 476622

In vivo Characterization
Methods used to characterize site in vivo:	mass spectrometry ( 1 , 2 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 12 , 13 , 14 , 15 , 16 , 17 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 ) , phospho-antibody ( 41 ) , western blotting ( 41 )
Disease tissue studied:	breast cancer ( 7 , 14 ) , breast ductal carcinoma ( 7 ) , HER2 positive breast cancer ( 2 ) , luminal A breast cancer ( 2 ) , luminal B breast cancer ( 2 ) , breast cancer, surrounding tissue ( 2 ) , breast cancer, triple negative ( 2 , 7 ) , cervical cancer ( 39 ) , cervical adenocarcinoma ( 39 ) , leukemia ( 19 ) , acute myelogenous leukemia ( 19 ) , lung cancer ( 9 , 15 ) , non-small cell lung cancer ( 15 ) , non-small cell lung adenocarcinoma ( 9 ) , neuroblastoma ( 13 ) , ovarian cancer ( 7 ) , melanoma skin cancer ( 4 )
Relevant cell line - cell type - tissue:	293 (epithelial) ( 6 , 43 ) , 293E (epithelial) ( 28 ) , breast ( 2 , 7 ) , DG75 (B lymphocyte) ( 32 ) , Flp-In T-Rex-293 (epithelial) [PRKD1 (human), genetic knockin] ( 17 ) , Flp-In T-Rex-293 (epithelial) ( 17 ) , H2077 (pulmonary) ( 15 ) , H322M (pulmonary) ( 15 ) , HCC366 (pulmonary) ( 15 ) , HCC78 (pulmonary) ( 15 ) , HEK293T (epithelial) ( 49 ) , HeLa (cervical) ( 1 , 5 , 12 , 31 , 41 , 45 , 47 , 50 ) , HeLa S3 (cervical) ( 39 ) , HUES-7 ('stem, embryonic') ( 42 ) , HUES-9 ('stem, embryonic') ( 30 ) , Jurkat (T lymphocyte) ( 10 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 29 , 33 , 34 , 35 , 36 , 37 , 38 , 40 , 44 , 46 , 48 ) , K562 (erythroid) ( 12 ) , KG-1 (myeloid) ( 19 ) , lung ( 9 ) , NB10 (neural crest) ( 13 ) , NPC (neural crest) ( 13 ) , ovary ( 7 ) , PANC-1 (pancreatic) [PRP4 (human), knockdown, Lentiviral introduced doxycycline-inducible PRP4 shRNA] ( 8 ) , PANC-1 (pancreatic) ( 8 ) , SKBr3 (breast cell) ( 14 ) , T lymphocyte-blood ( 16 ) , WM239A (melanocyte) ( 4 )

Upstream Regulation
Regulatory protein:	PRP4 (human) ( 8 )
Putative in vivo kinases:	CDK1 (human) ( 41 ) , ERK1 (human) ( 41 ) , ERK2 (human) ( 41 )
Kinases, in vitro:	CDK1 (human) ( 41 ) , ERK1 (human) ( 41 ) , ERK2 (human) ( 41 )
Treatments:	antibody ( 16 , 46 ) , EGF ( 41 ) , ischemia ( 7 ) , nocodazole ( 39 ) , phorbol_ester ( 41 ) , selumetinib ( 4 ) , U0126 ( 41 ) , vemurafenib ( 4 )

Downstream Regulation
Effects of modification on NUP50:	molecular association, regulation ( 41 )
Inhibit interaction with:	KPNB1 (human) ( 41 )

References
1	Huang H, et al. (2016) Simultaneous Enrichment of Cysteine-containing Peptides and Phosphopeptides Using a Cysteine-specific Phosphonate Adaptable Tag (CysPAT) in Combination with titanium dioxide (TiO2) Chromatography. Mol Cell Proteomics 15, 3282-3296 27281782 Curated Info
2	Mertins P, et al. (2016) Proteogenomics connects somatic mutations to signalling in breast cancer. Nature 534, 55-62 27251275 Curated Info
3	Boeing S, et al. (2016) Multiomic Analysis of the UV-Induced DNA Damage Response. Cell Rep 15, 1597-1610 27184836 Curated Info
4	Stuart SA, et al. (2015) A Phosphoproteomic Comparison of B-RAFV600E and MKK1/2 Inhibitors in Melanoma Cells. Mol Cell Proteomics 14, 1599-615 25850435 Curated Info
5	Sharma K, et al. (2014) Ultradeep human phosphoproteome reveals a distinct regulatory nature of Tyr and Ser/Thr-based signaling. Cell Rep 8, 1583-94 25159151 Curated Info
6	Wang R, et al. (2014) Global discovery of high-NaCl-induced changes of protein phosphorylation. Am J Physiol Cell Physiol 307, C442-54 24965592 Curated Info
7	Mertins P, et al. (2014) Ischemia in tumors induces early and sustained phosphorylation changes in stress kinase pathways but does not affect global protein levels. Mol Cell Proteomics 13, 1690-704 24719451 Curated Info
8	Gao Q, et al. (2013) Evaluation of cancer dependence and druggability of PRP4 kinase using cellular, biochemical, and structural approaches. J Biol Chem 288, 30125-38 24003220 Curated Info
9	Schweppe DK, Rigas JR, Gerber SA (2013) Quantitative phosphoproteomic profiling of human non-small cell lung cancer tumors. J Proteomics 91, 286-96 23911959 Curated Info
10	Mertins P, et al. (2013) Integrated proteomic analysis of post-translational modifications by serial enrichment. Nat Methods 10, 634-7 23749302 Curated Info
11	Shiromizu T, et al. (2013) Identification of missing proteins in the neXtProt database and unregistered phosphopeptides in the PhosphoSitePlus database as part of the Chromosome-centric Human Proteome Project. J Proteome Res 12, 2414-21 23312004 Curated Info
12	Zhou H, et al. (2013) Toward a comprehensive characterization of a human cancer cell phosphoproteome. J Proteome Res 12, 260-71 23186163 Curated Info
13	DeNardo BD, et al. (2013) Quantitative phosphoproteomic analysis identifies activation of the RET and IGF-1R/IR signaling pathways in neuroblastoma. PLoS One 8, e82513 24349301 Curated Info
14	Imami K, et al. (2012) Temporal profiling of lapatinib-suppressed phosphorylation signals in EGFR/HER2 pathways. Mol Cell Proteomics 11, 1741-57 22964224 Curated Info
15	Klammer M, et al. (2012) Phosphosignature predicts dasatinib response in non-small cell lung cancer. Mol Cell Proteomics 11, 651-68 22617229 Curated Info
16	Ruperez P, Gago-Martinez A, Burlingame AL, Oses-Prieto JA (2012) Quantitative phosphoproteomic analysis reveals a role for serine and threonine kinases in the cytoskeletal reorganization in early T cell receptor activation in human primary T cells. Mol Cell Proteomics 11, 171-86 22499768 Curated Info
17	Franz-Wachtel M, et al. (2012) Global detection of protein kinase D-dependent phosphorylation events in nocodazole-treated human cells. Mol Cell Proteomics 11, 160-70 22496350 Curated Info
18	Beli P, et al. (2012) Proteomic Investigations Reveal a Role for RNA Processing Factor THRAP3 in the DNA Damage Response. Mol Cell 46, 212-25 22424773 Curated Info
19	Weber C, Schreiber TB, Daub H (2012) Dual phosphoproteomics and chemical proteomics analysis of erlotinib and gefitinib interference in acute myeloid leukemia cells. J Proteomics 75, 1343-56 22115753 Curated Info
20	Mulhern D (2011) CST Curation Set: 12699; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
21	Mulhern D (2011) CST Curation Set: 12700; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
22	Mulhern D (2011) CST Curation Set: 12701; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
23	Mulhern D (2011) CST Curation Set: 12702; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
24	Mulhern D (2011) CST Curation Set: 12704; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: DpSG Curated Info
25	Mulhern D (2011) CST Curation Set: 12572; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
26	Mulhern D (2011) CST Curation Set: 12569; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
27	Mulhern D (2011) CST Curation Set: 12571; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
28	Hsu PP, et al. (2011) The mTOR-regulated phosphoproteome reveals a mechanism of mTORC1-mediated inhibition of growth factor signaling. Science 332, 1317-22 21659604 Curated Info
29	Guo A (2011) CST Curation Set: 11288; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-Thr-Pro Motif Antibody (polyAB) Cat#: 3003, PTMScan(R) Phospho-Thr-Pro Motif (T*P) Immunoaffinity Beads Cat#: 1996 Curated Info
30	Rigbolt KT, et al. (2011) System-wide temporal characterization of the proteome and phosphoproteome of human embryonic stem cell differentiation. Sci Signal 4, rs3 21406692 Curated Info
31	Hegemann B, et al. (2011) Systematic phosphorylation analysis of human mitotic protein complexes. Sci Signal 4, rs12 22067460 Curated Info
32	Iliuk AB, et al. (2010) In-depth analyses of kinase-dependent tyrosine phosphoproteomes based on metal ion-functionalized soluble nanopolymers. Mol Cell Proteomics 9, 2162-72 20562096 Curated Info
33	Possemato A (2010) CST Curation Set: 9793; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSP Curated Info
34	Possemato A (2010) CST Curation Set: 9640; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
35	Possemato A (2010) CST Curation Set: 9646; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
36	Possemato A (2010) CST Curation Set: 9645; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
37	Possemato A (2010) CST Curation Set: 9642; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
38	Possemato A (2010) CST Curation Set: 8993; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-Thr-Pro Motif Antibody (polyAB) Cat#: 3003, PTMScan(R) Phospho-Thr-Pro Motif (T*P) Immunoaffinity Beads Cat#: 1996 Curated Info
39	Olsen JV, et al. (2010) Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis. Sci Signal 3, ra3 20068231 Curated Info
40	Possemato A (2009) CST Curation Set: 8351; Year: 2009; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (K/R)XX[ST] Curated Info
41	Kosako H, et al. (2009) Phosphoproteomics reveals new ERK MAP kinase targets and links ERK to nucleoporin-mediated nuclear transport. Nat Struct Mol Biol 16, 1026-35 19767751 Curated Info
42	Van Hoof D, et al. (2009) Phosphorylation dynamics during early differentiation of human embryonic stem cells. Cell Stem Cell 5, 214-26 19664995 Curated Info
43	Gauci S, et al. (2009) Lys-N and trypsin cover complementary parts of the phosphoproteome in a refined SCX-based approach. Anal Chem 81, 4493-501 19413330 Curated Info
44	Possemato A (2009) CST Curation Set: 6371; Year: 2009; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSP Curated Info
45	Chen RQ, et al. (2009) CDC25B mediates rapamycin-induced oncogenic responses in cancer cells. Cancer Res 69, 2663-8 19276368 Curated Info
46	Mayya V, et al. (2009) Quantitative phosphoproteomic analysis of T cell receptor signaling reveals system-wide modulation of protein-protein interactions. Sci Signal 2, ra46 19690332 Curated Info
47	Dephoure N, et al. (2008) A quantitative atlas of mitotic phosphorylation. Proc Natl Acad Sci U S A 105, 10762-7 18669648 Curated Info
48	Possemato A (2008) CST Curation Set: 4737; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (K/R)XX[ST] Curated Info
49	Molina H, et al. (2007) Global proteomic profiling of phosphopeptides using electron transfer dissociation tandem mass spectrometry. Proc Natl Acad Sci U S A 104, 2199-204 17287340 Curated Info
50	Beausoleil SA, et al. (2006) A probability-based approach for high-throughput protein phosphorylation analysis and site localization. Nat Biotechnol 24, 1285-92 16964243 Curated Info