Curated Information
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Home > Curated Information Page > PubMed Id: 24764294
Liu S, Premont RT, Rockey DC (2014) Endothelial nitric-oxide synthase (eNOS) is activated through G-protein-coupled receptor kinase-interacting protein 1 (GIT1) tyrosine phosphorylation and Src protein. J Biol Chem 289, 18163-74 24764294
This page summarizes selected information from the record referenced above and curated into PhosphoSitePlus®, a comprehensive online resource for the study of protein post-translational modifications (NAR, 2015, 43:D512-20). To learn more about the scope of PhosphoSitePlus®, click here.
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S473-p - Akt1 (rat)
Modsite: RPHFPQFsYSASGTA SwissProt Entrez-Gene
Orthologous residues
Akt1 (human): S473‑p, Akt1 iso2 (human): S411‑p, Akt1 (mouse): S473‑p, Akt1 (rat): S473‑p, Akt1 (fruit fly): S586‑p, Akt1 (cow): S473‑p
Characterization
Methods used to characterize site in vivo phospho-antibody, western blotting
Relevant cell lines - cell types - tissues:  endothelial-liver
Cellular systems studied:  primary cultured cells
Species studied:  rat
Comments:  liver sinusoids

S1176-p - eNOS (rat)
Modsite: TSRIRtQsFsLQERQ SwissProt Entrez-Gene
Orthologous residues
eNOS (human): S1177‑p, eNOS (mouse): S1176‑p, eNOS (rat): S1176‑p, eNOS (rabbit): S1183‑p, eNOS (pig): S1179‑p, eNOS (cow): S1179‑p
Characterization
Methods used to characterize site in vivo phospho-antibody, western blotting
Relevant cell lines - cell types - tissues:  endothelial-liver
Cellular systems studied:  primary cultured cells
Species studied:  rat
Comments:  liver sinusoids
Upstream Regulation
Potential in vivo enzymes for site: 
Type Enzyme Evidence Notes
KINASE Akt1 (human) pharmacological inhibitor of upstream enzyme, transfection of constitutively active enzyme, phospho-antibody, transfection of dominant-negative enzyme
Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
GIT1 (rat) increase
endothelin increase
PP2 endothelin inhibit treatment-induced increase
PTX endothelin no effect upon treatment-induced increase
GRK2 (rat) increase GRK2 c-terminal
endothelin increase
PP2 endothelin inhibit treatment-induced increase
endothelin increase
PP2 endothelin inhibit treatment-induced increase

Y293-p - GIT1 (rat)
Modsite: EELAMDVyDEVDRRE SwissProt Entrez-Gene
Orthologous residues
GIT1 (human): Y284‑p, GIT1 iso3 (human): Y293‑p, GIT1 (mouse): Y293‑p, GIT1 (rat): Y293‑p
Characterization
Methods used to characterize site in vivo immunoprecipitation, mutation of modification site, phospho-antibody, western blotting
Relevant cell lines - cell types - tissues:  endothelial-liver
Cellular systems studied:  primary cultured cells
Species studied:  rat
Comments:  liver sinusoids
Upstream Regulation
Potential in vivo enzymes for site: 
Type Enzyme Evidence Notes
KINASE Src (rat) transfection of wild-type enzyme, transfection of inactive enzyme
Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
endothelin increase
endothelin GRK2 (rat) inhibit treatment-induced increase
PTX endothelin 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
eNOS (rat) Induces pull-down assay

Y554-p - GIT1 (rat)
Modsite: ELEDDAIySVHVPAG SwissProt Entrez-Gene
Orthologous residues
GIT1 (human): Y545‑p, GIT1 iso3 (human): Y554‑p, GIT1 (mouse): Y554‑p, GIT1 (rat): Y554‑p
Characterization
Methods used to characterize site in vivo immunoprecipitation, mutation of modification site, phospho-antibody, western blotting
Relevant cell lines - cell types - tissues:  endothelial-liver
Cellular systems studied:  primary cultured cells
Species studied:  rat
Comments:  liver sinusoids
Upstream Regulation
Potential in vivo enzymes for site: 
Type Enzyme Evidence Notes
KINASE Src (rat) transfection of wild-type enzyme, transfection of inactive enzyme
Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
endothelin increase
endothelin GRK2 (rat) inhibit treatment-induced increase
PTX endothelin 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
eNOS (rat) Induces pull-down assay

Y419-p - Src (rat)
Modsite: RLIEDNEyTARQGAk SwissProt Entrez-Gene
Orthologous residues
Src (human): Y419‑p, Src iso2 (human): Y425‑p, Src (mouse): Y424‑p, Src iso2 (mouse): Y418‑p, Src (rat): Y419‑p, Src (chicken): Y416‑p
Characterization
Methods used to characterize site in vivo phospho-antibody, western blotting
Relevant cell lines - cell types - tissues:  endothelial-liver
Cellular systems studied:  primary cultured cells
Species studied:  rat
Comments:  liver sinusoids
Upstream Regulation
Treatments, proteins and their effect on site modification: 
Treatments Referenced Treatments Manipulated Protein Referenced Protein Effect Notes
GIT1 (rat) increase
endothelin increase
PP2 endothelin inhibit treatment-induced increase
PTX endothelin no effect upon treatment-induced increase
GRK2 (rat) increase GRK2 c-terminal