Direct cysteine sulfenylation drives activation of the Src kinase

Heppner, David E.; Dustin, Christopher M.; Liao, Chenyi; Hristova, Milena; Veith, Carmen; Little, Andrew C.; Ahlers, Bethany A.; White, Sheryl L.; Deng, Bin; Lam, Ying‐Wai; Li, Jianing; Vliet, Albert van der

doi:10.1038/s41467-018-06790-1

Cited by 100 publications

(92 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…9B). Notably, both SRC and FGFR have previously been shown to be redox-regulated through Cys oxidation mechanisms in the P-loop (53,54). However, whether these kinases also form a strand exchange dimer similar to AtFN3K is not known, and it will be interesting to attempt to solve the structure of Tyr kinases crystallized under different oxidizing conditions.…”

Section: Discussionmentioning

confidence: 99%

A novel redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase superfamily

Shrestha

Katiyar

Sanz-Rodriguez

et al. 2020

Preprint

View full text Add to dashboard Cite

words)Aberrant regulation of metabolic kinases by altered redox homeostasis is a major contributing factor in aging and disease such as diabetes. However, the biochemical mechanisms by which metabolic kinases are regulated under oxidative stress is poorly understood. In this study, we demonstrate that the catalytic activity of a conserved family of Fructosamine-3-kinases (FN3Ks), which are evolutionarily related to eukaryotic protein kinases (ePKs), are regulated by redox active cysteines in the kinase domain. By solving the crystal structure of FN3K homolog from Arabidopsis thaliana (AtFN3K), we demonstrate that it forms an unexpected strandexchange dimer in which the ATP binding P-loop and adjoining beta strands are swapped between two chains in the dimer. This dimeric configuration is characterized by strained inter-chain disulfide bonds that stabilize the P-loop in an extended conformation and tethers the substratebinding lobes in a unique inactive conformation. Mutational analysis and solution studies confirm that the strained disulfides function as redox "switches" to reversibly regulate FN3K activity and dimerization. Consistently, we find that human FN3K, which contains an equivalent P-loop Cys, is also redox sensitive, whereas ancestral bacterial FN3K homologs, which lack a P-loop Cys, are not. Furthermore, CRISPR knockout of FN3K in human HepG2 cells results in significant upregulation of redox metabolites including glutathione. We propose that redox regulation evolved progressively in FN3Ks in response to changing cellular redox conditions. Our studies provide important new insights into the origin and evolution of redox regulation in the protein kinase superfamily and open new avenues for targeting human FN3K in diabetic complications.

show abstract

Section: Discussionmentioning

confidence: 99%

A novel redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase superfamily

Shrestha

Katiyar

Sanz-Rodriguez

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Although these techniques have allowed identification of numerous human proteins undergoing Cys ox-PTMs in vivo, the report of specific oxidation sites by MS is very often limited to proteins highly abundant and/or highly susceptible to oxidation (13,14,67). In most cases, identification of the oxidized residue by MS relied on the production of recombinant proteins and in vitro oxidation (19)(20)(21). In the present study, we successfully used Mal-PEG 2 -Bio-based labeling coupled to InsPEx workflow to identify novel Cys ox-PTM sites in vivo by LC-MS/MS ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Pinpointing of cysteine oxidation sites in vivo by high-resolution proteomics reveals mechanism of redox-dependent inhibition of STING

Cuervo

Fortin

Caron

et al. 2020

Preprint

View full text Add to dashboard Cite

Brief summary of the main results.The function of proteins is regulated by post-translational modifications, among which reversible oxidation of Cys recently emerged as a key component. Comprehension of redox regulation of cellular responses requires identification of specific oxidation sites in vivo. Using a bioswitch method to specifically label Cys subjected to reversible oxidation coupled to mass spectrometry, we identified thousands of novel oxidation sites. Many are relevant to virus -host interaction pathways. Here, we focused on the oxid ation of STING, an adaptor critical for activating the innate immune type I interferon pathway engaged upon cytosolic DNA sensing. Molecular studies led us to establish a new model in which STING Cys 148 is oxidized at basal levels, while Cys 206 oxidation is induced by oxidative stress and ligand binding. We show that oxidation of Cys 206 has an inhibitory function to prevent STING hyperactivation. This study provides ground for novel research avenues aimed at designing therapeutics that target this pathway. AbstractProtein function is regulated by post-translational modifications, among which reversible oxidation of Cys (Cys ox-PTM) emerged as a key regulatory mechanism of cellular responses. The redox regulation of virus-host interactions is well documented, but in most cases, proteins subjected to Cys ox-PTM remain unknown. The identification of Cys ox-PTM sites in vivo is essential to underpin our understanding of the mechanisms of the redox regulation. In this study, we present a proteome-wide identification of reversible Cys ox-PTM sites in vivo during stimulation by oxidants using a maleimide-based bioswitch method coupled to mass spectrometry. We identified 2720 unique Cys ox-PTM sites encompassing 1473 proteins with distinct abundance, location and functions. Label-free quantification (LFQ)-based analysis revealed the enrichment of ox-PTM in numerous pathways, many relevant to virus-host interaction. Here, we focused on the oxidation of STING, the central adaptor of the innate immune type I interferon 2 pathway induced upon detection of cytosolic DNA. We provide the first in vivo demonstration of reversible oxidation of Cys 148 and Cys 206 of STING. Molecular analyses led us to establish a new model in which Cys 148 oxidation is constitutive, while Cys 206 oxidation is inducible by oxidative stress or by the natural ligand 2'3'-cGAMP. We show that oxidation of Cys 206 has an inhibitory function to prevent STING hyperactivation through the constraint of a conformational change associated with the formation of inactive polymers containing intermolecular disulfide bonds. This provides new ground for the design of therapies targeting STING relevant to autoinflammatory disorders, immunotherapies and vaccines.

show abstract

“…Two Src cysteine residues, Cys 185 and Cys 277, were identified as targets for H 2 O 2 ‐mediated sulfenylation (Cys‐SOH) in redox‐dependent kinase activation. The modification of these cysteines to Cys‐SOH induces local structural changes that directly impact the regulatory pTyr 527 and Tyr 416 residues …”

Section: Approaches For Ptm Identificationmentioning

confidence: 99%

Advances of Proteomics in Novel PTM Discovery: Applications in Cancer Therapy

Wang

Zhang

et al. 2019

Small Methods

View full text Add to dashboard Cite

Protein posttranslational modification (PTM) is a critical mechanism to enhance the diversity of protein species and functions in organisms. Currently, many different PTMs are discovered and found to be involved in a wide range of cellular processes. The aberrant regulation of PTM dynamics is reported to contribute to cancer development. Phosphorylation and acetylation are the best studied PTMs, and the application of high‐resolution mass spectrometry‐based proteomics and specific antibody‐based enrichment technologies significantly promotes novel PTM discovery. This review discusses methods for global PTM identification, including antibody‐based enrichment and chemical probe‐based identification. Several acylations, such as propionylation, butyrylation, succinylation, malonylation, and crotonylation, are recently found to influence both histone and nonhistone proteins and to be intimately linked with the progression of cancer. It is proposed that targeting novel PTM dynamics by pharmacological inhibitors has great potential for cancer therapy.

show abstract

Direct cysteine sulfenylation drives activation of the Src kinase

Cited by 100 publications

References 67 publications

A novel redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase superfamily

A novel redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase superfamily

Pinpointing of cysteine oxidation sites in vivo by high-resolution proteomics reveals mechanism of redox-dependent inhibition of STING

Advances of Proteomics in Novel PTM Discovery: Applications in Cancer Therapy

Contact Info

Product

Resources

About