Thr231

Site Information
tRNKVRktFVGtPCW SwissProt Entrez-Gene
Blast this site against: NCBI SwissProt PDB
Site Group ID: 456505
Available spectra: 3 CST

In vivo Characterization
Methods used to characterize site in vivo:	mass spectrometry ( 1 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 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 , 51 , 52 , 53 , 54 , 55 ) , mutation of modification site ( 19 , 57 ) , phospho-antibody ( 41 , 56 , 57 ) , western blotting ( 41 , 56 , 57 )
Disease tissue studied:	HER2 positive breast cancer ( 1 ) , luminal A breast cancer ( 1 ) , luminal B breast cancer ( 1 ) , breast cancer, triple negative ( 1 ) , gastric cancer ( 18 , 23 , 24 , 25 , 27 , 30 , 31 , 32 , 34 , 35 , 36 , 39 , 40 , 44 , 48 , 50 ) , gastric carcinoma ( 18 , 23 , 24 , 25 , 27 , 30 , 31 , 32 , 34 , 35 , 36 , 39 , 40 , 44 , 48 , 50 ) , lung cancer ( 3 , 4 , 5 , 6 , 8 , 26 , 28 , 29 , 33 , 37 , 38 , 42 , 45 , 47 , 49 ) , non-small cell lung cancer ( 4 , 6 , 7 , 8 , 26 , 28 , 29 , 33 , 37 , 38 , 42 , 45 , 47 , 49 ) , non-small cell lung adenocarcinoma ( 4 , 5 , 6 , 28 , 29 , 33 , 38 , 42 , 45 , 47 , 49 ) , non-small cell squamous cell lung carcinoma ( 5 , 28 , 29 , 33 , 38 , 45 , 47 , 49 ) , small-cell lung cancer ( 3 )
Relevant cell line - cell type - tissue:	'stem, embryonic' ( 19 ) , 293 (epithelial) ( 56 , 57 ) , A549 (pulmonary) ( 4 ) , breast ( 1 ) , Cal-12T (pulmonary) ( 6 ) , DMS79 (pulmonary) ( 3 ) , erythrocyte-blood ( 41 ) , H2009 (pulmonary) ( 8 ) , H2887 (pulmonary) ( 8 ) , HCC15 (pulmonary) ( 5 ) , HCC44 (pulmonary) ( 6 ) , HeLa (cervical) ( 46 , 51 , 52 , 53 , 54 , 55 , 57 ) , HOP62 (pulmonary) ( 8 ) , Jurkat (T lymphocyte) ( 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 20 , 21 , 22 , 43 ) , LOU-NH91 (squamous) ( 5 ) , lung ( 3 , 4 , 5 , 6 ) , MKN-45 (gastric) ( 9 , 18 , 23 , 24 , 25 , 27 , 30 , 31 , 32 , 34 , 35 , 36 , 39 , 40 , 44 , 48 , 50 ) , NCI-H128 (pulmonary) ( 3 ) , NCI-H1355 (pulmonary) ( 4 ) , NCI-H1417 (pulmonary) ( 3 ) , NCI-H1437 (pulmonary) ( 6 ) , NCI-H1666 (pulmonary) ( 6 ) , NCI-H1703 (squamous) ( 5 , 9 , 28 , 29 , 33 , 38 , 45 , 47 , 49 ) , NCI-H1792 (pulmonary) ( 4 ) , NCI-H2073 (pulmonary) ( 5 ) , NCI-H209 (pulmonary) ( 3 ) , NCI-H23 (pulmonary) ( 4 ) , NCI-H2342 (pulmonary) ( 5 ) , NCI-H2405 (pulmonary) ( 6 ) , NCI-H3122 (pulmonary) ( 7 ) , NCI-H322 (pulmonary) ( 8 ) , NCI-H3255 (pulmonary) ( 9 , 26 , 37 ) , NCI-H441 (pulmonary) ( 4 , 42 ) , NCI-H524 (pulmonary) ( 3 )

Upstream Regulation
Regulatory protein:	WNK1 (human) ( 41 , 57 )
Kinases, in vitro:	WNK1 (human) ( 58 )
Treatments:	A-769662 ( 41 ) , hypotonic_buffer ( 56 ) , sorbitol ( 57 ) , sucrose ( 41 )

Downstream Regulation
Effects of modification on STLK3:	activity, induced ( 19 ) , enzymatic activity, induced ( 56 , 57 ) , phosphorylation ( 41 )

References
1	Mertins P, et al. (2016) Proteogenomics connects somatic mutations to signalling in breast cancer. Nature 534, 55-62 27251275 Curated Info
2	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
3	Rikova K, Hall B (2013) CST Curation Set: 20734, 21161, 30112, 30153, 30154; Year: 2013; Biosample/Treatment: cell line, H1417, DMS79, H128, H209, H524; Disease: -; TMT: Y; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pY, p[ST], RXXp[ST], pSQ, p[ST]QG, LXRXXp[ST], p[ST]P Curated Info
4	Rikova K, Hall B (2013) CST Curation Set: 20736, 21163, 30158, 30159, 30160; Year: 2013; Biosample/Treatment: cell line, A549, H1355, H23, H441, H1792; Disease: -; TMT: Y; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pY, p[ST], RXXp[ST], pSQ, p[ST]QG, LXRXXp[ST], p[ST]P Curated Info
5	Rikova K, Hall B (2013) CST Curation Set: 20739, 21166, 30167, 30168, 30169; Year: 2013; Biosample/Treatment: cell line, H2342, H2073, Lou-NH91, HCC15, H1703; Disease: -; TMT: Y; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pY, p[ST], RXXp[ST], pSQ, p[ST]QG, LXRXXp[ST], p[ST]P Curated Info
6	Rikova K, Hall B (2013) CST Curation Set: 20741, 21168, 30173, 30174, 30175; Year: 2013; Biosample/Treatment: cell line, H1666, CAL-12T, H2405, HCC44, H1437; Disease: -; TMT: Y; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pY, p[ST], RXXp[ST], pSQ, p[ST]QG, LXRXXp[ST], p[ST]P Curated Info
7	Rikova K (2013) CST Curation Set: 18459; Year: 2013; Biosample/Treatment: cell line, H3122/crizotinib, geldanamycin; Disease: -; TMT: Y; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST](AGC) Curated Info
8	Klammer M, et al. (2012) Phosphosignature predicts dasatinib response in non-small cell lung cancer. Mol Cell Proteomics 11, 651-68 22617229 Curated Info
9	Stokes MP, et al. (2012) PTMScan Direct: Identification and Quantification of Peptides from Critical Signaling Proteins by Immunoaffinity Enrichment Coupled with LC-MS/MS. Mol Cell Proteomics 11, 187-201 22322096 Curated Info
10	Guo A (2012) CST Curation Set: 13982; Year: 2012; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
11	Mulhern D (2012) CST Curation Set: 13827; Year: 2012; 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)XpSX(K/R) Curated Info
12	Mulhern D (2012) CST Curation Set: 13828; Year: 2012; 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)XpSX(K/R) Curated Info
13	Mulhern D (2012) CST Curation Set: 13829; Year: 2012; 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)XpSX(K/R) Curated Info
14	Mulhern D (2012) CST Curation Set: 13830; Year: 2012; 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)XpSX(K/R) Curated Info
15	Mulhern D (2012) CST Curation Set: 13107; Year: 2012; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY]) Curated Info
16	Mulhern D (2012) CST Curation Set: 13108; Year: 2012; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY]) Curated Info
17	Mulhern D (2012) CST Curation Set: 13109; Year: 2012; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
18	Mulhern D (2012) CST Curation Set: 13405; Year: 2012; Biosample/Treatment: cell line, MKN-45/untreated; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
19	Thastrup JO, et al. (2012) SPAK/OSR1 regulate NKCC1 and WNK activity: analysis of WNK isoform interactions and activation by T-loop trans-autophosphorylation. Biochem J 441, 325-37 22032326 Curated Info
20	Mulhern D (2011) CST Curation Set: 13110; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Curated Info
21	Guo A (2011) CST Curation Set: 12018; 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 Curated Info
22	Guo A (2011) CST Curation Set: 11173; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Curated Info
23	Moritz A (2010) CST Curation Set: 10603; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
24	Moritz A (2010) CST Curation Set: 10602; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
25	Moritz A (2010) CST Curation Set: 10601; Year: 2010; Biosample/Treatment: cell line, MKN-45/DMSO; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
26	Moritz A (2010) CST Curation Set: 10599; Year: 2010; Biosample/Treatment: cell line, NCI-H3255/DMSO; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
27	Moritz A (2010) CST Curation Set: 10600; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
28	Moritz A (2010) CST Curation Set: 10595; Year: 2010; Biosample/Treatment: cell line, NCI-H1703/Su11274; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
29	Moritz A (2010) CST Curation Set: 10596; Year: 2010; Biosample/Treatment: cell line, NCI-H1703/DMSO; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
30	Moritz A (2010) CST Curation Set: 10233; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
31	Moritz A (2010) CST Curation Set: 10238; Year: 2010; Biosample/Treatment: cell line, MKN-45/DMSO; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
32	Moritz A (2010) CST Curation Set: 10235; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
33	Moritz A (2010) CST Curation Set: 10240; Year: 2010; Biosample/Treatment: cell line, NCI-H1703/DMSO; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
34	Moritz A (2010) CST Curation Set: 10232; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
35	Moritz A (2010) CST Curation Set: 10237; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
36	Moritz A (2010) CST Curation Set: 10234; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
37	Moritz A (2010) CST Curation Set: 10241; Year: 2010; Biosample/Treatment: cell line, NCI-H3255/Iressa; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
38	Moritz A (2010) CST Curation Set: 10239; Year: 2010; Biosample/Treatment: cell line, NCI-H1703/Gleevec; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
39	Moritz A (2010) CST Curation Set: 10236; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
40	Moritz A (2010) CST Curation Set: 10231; Year: 2010; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY] Curated Info
41	Sid B, et al. (2010) Stimulation of human and mouse erythrocyte Na(+)-K(+)-2Cl(-) cotransport by osmotic shrinkage does not involve AMP-activated protein kinase, but is associated with STE20/SPS1-related proline/alanine-rich kinase activation. J Physiol 588, 2315-28 20442269 Curated Info
42	Moritz A (2010) CST Curation Set: 9986; Year: 2010; Biosample/Treatment: cell line, NCI-H441/untreated; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY]) Curated Info
43	Possemato A (2010) CST Curation Set: 9918; 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 Curated Info
44	Moritz A (2010) CST Curation Set: 9234; Year: 2010; Biosample/Treatment: cell line, MKN-45/calyculin_A & pervanadate; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY]) Curated Info
45	Moritz A (2010) CST Curation Set: 9240; Year: 2010; Biosample/Treatment: cell line, NCI-H1703/untreated; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY]) Curated Info
46	Moritz A (2010) CST Curation Set: 9237; Year: 2010; Biosample/Treatment: cell line, HeLa/nocodazole; Disease: cervical adenocarcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[STY]) Curated Info
47	Possemato A (2009) CST Curation Set: 8409; Year: 2009; Biosample/Treatment: cell line, NCI-H1703/untreated; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
48	Possemato A (2009) CST Curation Set: 8412; Year: 2009; Biosample/Treatment: cell line, MKN-45/untreated; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
49	Possemato A (2009) CST Curation Set: 8410; Year: 2009; Biosample/Treatment: cell line, NCI-H1703/untreated; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
50	Possemato A (2009) CST Curation Set: 8411; Year: 2009; Biosample/Treatment: cell line, MKN-45/untreated; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
51	Zhou J (2009) CST Curation Set: 7600; Year: 2009; Biosample/Treatment: cell line, HeLa/untreated; Disease: cervical adenocarcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
52	Zhou J (2009) CST Curation Set: 7598; Year: 2009; Biosample/Treatment: cell line, HeLa/untreated; Disease: cervical adenocarcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
53	Zhou J (2009) CST Curation Set: 7597; Year: 2009; Biosample/Treatment: cell line, HeLa/untreated; Disease: cervical adenocarcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
54	Zhou J (2009) CST Curation Set: 7596; Year: 2009; Biosample/Treatment: cell line, HeLa/untreated; Disease: cervical adenocarcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
55	Zhou J (2009) CST Curation Set: 7595; Year: 2009; Biosample/Treatment: cell line, HeLa/untreated; Disease: cervical adenocarcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
56	Richardson C, et al. (2008) Activation of the thiazide-sensitive Na+-Cl- cotransporter by the WNK-regulated kinases SPAK and OSR1. J Cell Sci 121, 675-84 18270262 Curated Info
57	Zagórska A, et al. (2007) Regulation of activity and localization of the WNK1 protein kinase by hyperosmotic stress. J Cell Biol 176, 89-100 17190791 Curated Info
58	Vitari AC, Deak M, Morrice NA, Alessi DR (2005) The WNK1 and WNK4 protein kinases that are mutated in Gordon's hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases. Biochem J 391, 17-24 16083423 Curated Info