Novel autophosphorylation sites of Src family kinases regulate kinase activity and <scp>SH</scp>2 domain‐binding capacity

Weir, Marion E.; Mann, Jacqueline E.; Corwin, Thomas; Fulton, Zachary W.; Hao, Jennifer M.; Maniscalco, Jeanine F.; Kenney, Marie; Roque, Kristal M. Roman; Chapdelaine, Elizabeth; Stelzl, Ulrich; Deming, Paula B.; Ballif, Bryan A.; Hinkle, Karen L.

doi:10.1002/1873-3468.12144

Cited by 13 publications

(13 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The phosphorylated amino acid residue (Ser206) localizes in the SH2 domain of Fgr; phosphorylation of which may modulates the binding affinity and specificity for SFKs (68,69). Ser 460 of HCK localizes in the kinase domain of this kinase; Tyrosine residue (Tyr316) of Lyn also localizes in the kinase domain of this protein which is equivalent to Tyr338 of Src and phosphorylation of this site was reported autophosphorylation by Src,(70) the phosphorylation of the domain is also reported autophosphorylated by SFKs, (69), and was predicted to be dominated by SFKs themselves in our analysis. The phosphorylation of Ser17 of Src, which is a specific residue in the N terminus of Src, will be discussed in greater detail in the following section.…”

Section: Phosphoproteome Profiles Of Hcc Tumor and Wild Typementioning

confidence: 99%

In Vivo Phosphoproteome Analysis Reveals Kinome Reprogramming in Hepatocellular Carcinoma

Lang

Wang³

et al. 2018

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Aberrant kinases contribute to cancer survival and proliferation. Here, we quantitatively characterized phosphoproteomic changes in an HBx-transgenic mouse model of hepatocellular carcinoma (HCC) using high-resolution mass spectrometry, profiled 22,539 phosphorylation sites on 5431 proteins. Using a strategy to interpret kinase- substrate relations in HCC and to uncover predominant kinases in tumors, our results, revealed elevated kinase activities of Src family kinases (SFKs), PKCs, MAPKs, and ROCK2 in HCC, representatives of which were further validated in cell models and clinical HBV-positive HCC samples. Inhibitor combinations targeting Src and PKCs or ROCK2 both synergized significantly to inhibit cell growth. In addition, we demonstrated that phosphorylation at Src Ser17 directly affects its kinase activity. Our phosphoproteome data facilitated the construction of a detailed molecular landscape in HCC and should serve as a resource for the cancer community. Our strategy is generally applicable to targeted therapeutics, also highlights potential mechanisms of kinase regulation.

show abstract

Section: Phosphoproteome Profiles Of Hcc Tumor and Wild Typementioning

confidence: 99%

In Vivo Phosphoproteome Analysis Reveals Kinome Reprogramming in Hepatocellular Carcinoma

Lang

Wang³

et al. 2018

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…Subsequent (auto)phosphorylation of Tyr-416 by intermolecular encounter with another active kinase then stabilizes the active form of Src 11 , 12 . In addition to Tyr-416, phosphorylation of additional tyrosines may also regulate SFK function 13 .…”

Section: Introductionmentioning

confidence: 99%

Direct cysteine sulfenylation drives activation of the Src kinase

et al. 2018

View full text Add to dashboard Cite

The Src kinase controls aspects of cell biology and its activity is regulated by intramolecular structural changes induced by protein interactions and tyrosine phosphorylation. Recent studies indicate that Src is additionally regulated by redox-dependent mechanisms, involving oxidative modification(s) of cysteines within the Src protein, although the nature and molecular-level impact of Src cysteine oxidation are unknown. Using a combination of biochemical and cell-based studies, we establish the critical importance of two Src cysteine residues, Cys-185 and Cys-277, as targets for H2O2-mediated sulfenylation (Cys-SOH) in redox-dependent kinase activation in response to NADPH oxidase-dependent signaling. Molecular dynamics and metadynamics simulations reveal the structural impact of sulfenylation of these cysteines, indicating that Cys-277-SOH enables solvent exposure of Tyr-416 to promote its (auto)phosphorylation, and that Cys-185-SOH destabilizes pTyr-527 binding to the SH2 domain. These redox-dependent Src activation mechanisms offer opportunities for development of Src-selective inhibitors in treatment of diseases where Src is aberrantly activated.

show abstract

“…The pFlag-CMV2 LARP4 and pFLAG-CMV2 plasmids were a gift from Richard Maraia from the National Institute of Health, previously described [24]. The Fyn-Y3D mutant (Fyn-Y3D), possessing three tyrosine-to-aspartate mutations in the SH2 domain (Y185D, Y213D, and Y214D), was described previously [25].…”

Section: Methodsmentioning

confidence: 99%

Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A

et al. 2018

View full text Add to dashboard Cite

The cAMP-dependent protein kinase A (PKA) is a serine/threonine kinase involved in many fundamental cellular processes, including migration and proliferation. Recently, we found that the Src family kinase Fyn phosphorylates the catalytic subunit of PKA (PKA-C) at Y69, thereby increasing PKA kinase activity. We also showed that Fyn induced the phosphorylation of cellular proteins within the PKA preferred target motif. This led to the hypothesis that Fyn could affect proteins in complex with PKA. To test this, we employed a quantitative mass spectrometry approach to identify Fyn-dependent binding partners in complex with PKA-C. We found Fyn enhanced the binding of PKA-C to several cytoskeletal regulators that localize to the centrosome and Golgi apparatus. Three of these Fyn-induced PKA interactors, AKAP9, PDE4DIP, and CDK5RAP2, were validated biochemically and were shown to exist in complex with Fyn and PKA in a glioblastoma cell line. Intriguingly, the complexes formed between PKA-C and these known AKAPs were dependent upon Fyn catalytic activity and expression levels. In addition, we identified Fyn-regulated phosphorylation sites on proteins in complex with PKA-C. We also identified and biochemically validated a novel PKA-C interactor, LARP4, which complexed with PKA in the absence of Fyn. These results demonstrate the ability of Fyn to influence the docking of PKA to specific cellular scaffolds and suggest that Fyn may affect the downstream substrates targeted by PKA.

show abstract

Novel autophosphorylation sites of Src family kinases regulate kinase activity and SH2 domain‐binding capacity

Cited by 13 publications

References 45 publications

In Vivo Phosphoproteome Analysis Reveals Kinome Reprogramming in Hepatocellular Carcinoma

In Vivo Phosphoproteome Analysis Reveals Kinome Reprogramming in Hepatocellular Carcinoma

Direct cysteine sulfenylation drives activation of the Src kinase

Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A

Contact Info

Product

Resources

About