Curated Information
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Curated Information Page
PubMed Id: 19917253 
This page summarizes selected information from the article referenced above and curated into PhosphoSitePlus®, a comprehensive online resource for the study of protein post-translational modifications (NAR, 2012,40:D261-70). To learn more about the scope of PhosphoSitePlus®, click here.
Gao S, et al. (2009) Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta signaling. Mol Cell 36, 457-68 19917253
Only sites from this record are displayed on this page. Click on the protein name to open the protein page, and on the RSD number to open the site page. For the complete dataset, click the download button, on the right.
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S342-p - NEDD4L (human) ▲
Orthologous residues
NEDD4L (human): S342‑p, NEDD4L iso5 (human): S342‑p, NEDD4L (mouse): S371‑p, NEDD4L iso3 (mouse): S371‑p, NEDD4L (rat): S330‑p
Characterization
 Methods used to characterize site in vivo:  [32P] bio-synthetic labeling, mutation of modification site, western blotting
 Relevant cell lines - cell types - tissues:  293T (epithelial)
 Cellular systems studied:  cell lines
 Species studied:  human
 Enzymes shown to modify site in vitro: 
Type Enzyme
KINASE SGK1 (human)
Downstream Regulation
 Effect of modification (function):  molecular association, regulation
 Modification regulates interactions with: 
Interacting molecule Interacting domains Effect Consequences (function) Consequences (process) Detection assays
Smad3 (human) Disrupts ubiquitination in vitro

S448-p - NEDD4L (human) ▲
Orthologous residues
NEDD4L (human): S448‑p, NEDD4L iso5 (human): S428‑p, NEDD4L (mouse): S477‑p, NEDD4L iso3 (mouse): S457‑p, NEDD4L (rat): S436‑p
Characterization
 Methods used to characterize site in vivo:  [32P] bio-synthetic labeling, mutation of modification site, western blotting
 Relevant cell lines - cell types - tissues:  293T (epithelial)
 Cellular systems studied:  cell lines
 Species studied:  human
 Enzymes shown to modify site in vitro: 
Type Enzyme
KINASE SGK1 (human)
Downstream Regulation
 Effect of modification (function):  molecular association, regulation
 Modification regulates interactions with: 
Interacting molecule Interacting domains Effect Consequences (function) Consequences (process) Detection assays
Smad3 (human) Disrupts ubiquitination in vitro

T220-p - Smad2 (human) ▲
Orthologous residues
Smad2 (human): T220‑p, Smad2 (mouse): T220‑p, Smad2 (rat): T220‑p
Characterization
 Methods used to characterize site in vivo:  mutation of modification site, phospho-antibody, western blotting
 Relevant cell lines - cell types - tissues:  HaCaT (keratinocyte)
 Cellular systems studied:  cell lines
 Species studied:  human
Upstream Regulation
 Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
TGF-beta increase
EGF increase

T179-p - Smad3 (human) ▲
Orthologous residues
Smad3 (human): T179‑p, Smad3 (mouse): T179‑p, Smad3 (rat): T179‑p
Characterization
 Methods used to characterize site in vivo:  mutation of modification site, phospho-antibody, western blotting
 Relevant cell lines - cell types - tissues:  293T (epithelial), HaCaT (keratinocyte)
 Cellular systems studied:  cell lines
 Species studied:  human
Upstream Regulation
 Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
EGF increase
TGF-beta increase
SB431542 TGF-beta inhibit treatment-induced increase
flavopiridol TGF-beta inhibit treatment-induced increase
U0126 TGF-beta no effect upon treatment-induced increase
Downstream Regulation
 Effect of modification (function):  molecular association, regulation
 Modification regulates interactions with: 
Interacting molecule Interacting domains Effect Consequences (function) Consequences (process) Detection assays
NEDD4L (human) Induces co-immunoprecipitation

S204-p - Smad3 (human) ▲
Orthologous residues
Smad3 (human): S204‑p, Smad3 (mouse): S204‑p, Smad3 (rat): S204‑p
Characterization
 Methods used to characterize site in vivo:  mutation of modification site, phospho-antibody, western blotting
 Relevant cell lines - cell types - tissues:  HaCaT (keratinocyte)
 Cellular systems studied:  cell lines
 Species studied:  human
Upstream Regulation
 Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
TGF-beta increase
EGF increase

S208-p - Smad3 (human) ▲
Orthologous residues
Smad3 (human): S208‑p, Smad3 (mouse): S208‑p, Smad3 (rat): S208‑p
Characterization
 Methods used to characterize site in vivo:  mutation of modification site, phospho-antibody, western blotting
 Relevant cell lines - cell types - tissues:  HaCaT (keratinocyte)
 Cellular systems studied:  cell lines
 Species studied:  human
Upstream Regulation
 Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
EGF increase
TGF-beta increase
SB431542 TGF-beta inhibit treatment-induced increase
flavopiridol TGF-beta inhibit treatment-induced increase
U0126 TGF-beta no effect upon treatment-induced increase

S213-p - Smad3 (human) ▲
Orthologous residues
Smad3 (human): S213‑p, Smad3 (mouse): S213‑p, Smad3 (rat): S213‑p
Characterization
 Methods used to characterize site in vivo:  mutation of modification site, phospho-antibody, western blotting
 Relevant cell lines - cell types - tissues:  HaCaT (keratinocyte)
 Cellular systems studied:  cell lines
 Species studied:  human
Upstream Regulation
 Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
TGF-beta increase
EGF increase


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