title: Phosphotyrosine Substrate Sequence Motifs for Dual Specificity Phosphatases.
abstract: Protein tyrosine phosphatases dephosphorylate tyrosine residues of proteins, whereas, dual specificity phosphatases (DUSPs) are a subgroup of protein tyrosine phosphatases that dephosphorylate not only Tyr(P) residue, but also the Ser(P) and Thr(P) residues of proteins. The DUSPs are linked to the regulation of many cellular functions and signaling pathways. Though many cellular targets of DUSPs are known, the relationship between catalytic activity and substrate specificity is poorly defined. We investigated the interactions of peptide substrates with select DUSPs of four types: MAP kinases (DUSP1 and DUSP7), atypical (DUSP3, DUSP14, DUSP22 and DUSP27), viral (variola VH1), and Cdc25 (A-C). Phosphatase recognition sites were experimentally determined by measuring dephosphorylation of 6,218 microarrayed Tyr(P) peptides representing confirmed and theoretical phosphorylation motifs from the cellular proteome. A broad continuum of dephosphorylation was observed across the microarrayed peptide substrates for all phosphatases, suggesting a complex relationship between substrate sequence recognition and optimal activity. Further analysis of peptide dephosphorylation by hierarchical clustering indicated that DUSPs could be organized by substrate sequence motifs, and peptide-specificities by phylogenetic relationships among the catalytic domains. The most highly dephosphorylated peptides represented proteins from 29 cell-signaling pathways, greatly expanding the list of potential targets of DUSPs. These newly identified DUSP substrates will be important for examining structure-activity relationships with physiologically relevant targets.
authors: Molecular and Translational Sciences Division, U.S. Army Medical Research Institute of Infectious Diseases, Frederick, MD, 21702, United States of America; The Geneva Foundation, Tacoma, WA, 98402, United States of America.
authors: Bryan M
authors: BM
authors: Zhao
authors: Molecular and Translational Sciences Division, U.S. Army Medical Research Institute of Infectious Diseases, Frederick, MD, 21702, United States of America.
authors: Sarah L
authors: SL
authors: Keasey
authors: Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, Maryland, 21702, United States of America.
authors: Joseph E
authors: JE
authors: Tropea
authors: Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, Maryland, 21702, United States of America; Basic Science Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, 21702, United States of America.
authors: George T
authors: GT
authors: Lountos
authors: Molecular and Translational Sciences Division, U.S. Army Medical Research Institute of Infectious Diseases, Frederick, MD, 21702, United States of America.
authors: Beverly K
authors: BK
authors: Dyas
authors: Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, Maryland, 21702, United States of America.
authors: Scott
authors: S
authors: Cherry
authors: Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, Maryland, 21702, United States of America.
authors: Sreejith
authors: S
authors: Raran-Kurussi
authors: Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, Maryland, 21702, United States of America.
authors: David S
authors: DS
authors: Waugh
authors: Molecular and Translational Sciences Division, U.S. Army Medical Research Institute of Infectious Diseases, Frederick, MD, 21702, United States of America.
authors: Robert G
authors: RG
authors: Ulrich
mesh_terms: D020816:Amino Acid Motifs; D054638:Dual Specificity Phosphatase 1 / Q000235:genetics / Q000378:metabolism; D054640:Dual Specificity Phosphatase 3 / Q000235:genetics / Q000378:metabolism; D054637:Dual-Specificity Phosphatases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D054639:Mitogen-Activated Protein Kinase Phosphatases / Q000235:genetics / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism*; D010802:Phylogeny; D040081:Protein Array Analysis; D011994:Recombinant Proteins; D015398:Signal Transduction; D013379:Substrate Specificity; D020687:cdc25 Phosphatases / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D019000:Phosphotyrosine; D054639:Mitogen-Activated Protein Kinase Phosphatases; D010749:Phosphoprotein Phosphatases; C522887:STYXL2 protein, human; C489855:dual specificity phosphatase 12; C517004:CDC25A protein, human; C517019:DUSP1 protein, human; C498843:DUSP14 protein, human; C442824:DUSP22 protein, human; C517025:DUSP3 protein, human; C107523:DUSP7 protein, human; D054638:Dual Specificity Phosphatase 1; D054640:Dual Specificity Phosphatase 3; D054637:Dual-Specificity Phosphatases; D020687:cdc25 Phosphatases