scholarly journals Spatial and temporal alterations in protein structure by EGF regulate cryptic cysteine oxidation

2019 ◽  
Author(s):  
Jessica B Behring ◽  
Sjoerd van der Post ◽  
Arshag D Mooradian ◽  
Matthew J Egan ◽  
Maxwell I Zimmerman ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Redox Regulation ◽  
Receptor Tyrosine Kinases ◽  
Redox Signaling ◽  
Signaling Networks ◽  
Cysteine Oxidation ◽  
Time Resolved ◽  
Cellular Redox ◽  
Redox Biology ◽  
Stimulation Of

AbstractStimulation of receptor tyrosine kinases (RTK) such as EGF locally increase reactive oxygen species (ROS) levels at the plasma membrane that oxidize cysteines in proteins to enhance downstream signaling. Spatial confinement of ROS is an important regulatory mechanism to redox signaling, but it remains unknown why stimulation of different receptor tyrosine kinases (RTKs) at the plasma membrane target distinct sets of downstream proteins. To uncover additional mechanisms specifying which cysteines are redox regulated by EGF stimulation, we performed time-resolved quantification of the oxidation of 4,200 cysteine sites subsequent to EGF stimulation in A431 cells. EGF induces three distinct spatiotemporal patterns of cysteine oxidation in functionally organized protein networks, consistent with the spatial confinement model. Unexpectedly, protein crystal structure analysis and molecular dynamic simulation indicate widespread redox regulation of cryptic cysteines that are only solvent exposed upon changes in protein conformation. Phosphorylation and increased flux of nucleotide substrates serve as two distinct modes by which EGF specifies which cryptic cysteines become solvent exposed and redox regulated. Since proteins structurally regulated by different RTKs or cellular perturbations are largely unique, solvent exposure and redox regulation of cryptic cysteines is an important mechanism contextually delineating redox signaling networks.Significance StatementCellular redox processes are interconnected, but are not in equilibrium. Thus, understanding the redox biology of cells requires a systems-level, rather than reductionist, approach. Factors specifying which cysteines are redox regulated by a stimulus remain poorly characterized but are critical to understanding the fundamental properties of redox signaling networks. Here, we show that EGF stimulation induces oxidation of specific cysteines in 3 distinct spatiotemporal patterns. Redox regulated proteins include many proteins in the EGF pathway as well as many cysteines with known functional importance. Many redox regulated cysteines are cryptic and solvent exposed by changes in protein structure that were induced by EGF treatment. The novel finding that cryptic cysteines are redox regulated has important implications for how redox signaling networks are specified and regulated to minimize crosstalk. In addition, this time-resolved dataset of the redox kinetics of 4,200 cysteine sites is an important resource for others and is an important technological achievement towards systems-level understanding of cellular redox biology.

scholarly journals Spatial and temporal alterations in protein structure by EGF regulate cryptic cysteine oxidation

2020 ◽  
Vol 13 (615) ◽  
pp. eaay7315 ◽  
Author(s):  
Jessica B. Behring ◽  
Sjoerd van der Post ◽  
Arshag D. Mooradian ◽  
Matthew J. Egan ◽  
Maxwell I. Zimmerman ◽  
...  
Keyword(s):  
Redox Regulation ◽  
Redox Signaling ◽  
Crystal Structure Analysis ◽  
Protein Crystal ◽  
Cysteine Residues ◽  
Cysteine Oxidation ◽  
Solvent Exposure ◽  
A431 Cells ◽  
Spatial Confinement ◽  
Stimulation Of

Stimulation of plasma membrane receptor tyrosine kinases (RTKs), such as the epidermal growth factor receptor (EGFR), locally increases the abundance of reactive oxygen species (ROS). These ROS then oxidize cysteine residues in proteins to potentiate downstream signaling. Spatial confinement of ROS is an important regulatory mechanism of redox signaling that enables the stimulation of different RTKs to oxidize distinct sets of downstream proteins. To uncover additional mechanisms that specify cysteines that are redox regulated by EGF stimulation, we performed time-resolved quantification of the EGF-dependent oxidation of 4200 cysteine sites in A431 cells. Fifty-one percent of cysteines were statistically significantly oxidized by EGF stimulation. Furthermore, EGF induced three distinct spatiotemporal patterns of cysteine oxidation in functionally organized protein networks, consistent with the spatial confinement model. Unexpectedly, protein crystal structure analysis and molecular dynamics simulations indicated widespread redox regulation of cryptic cysteine residues that are solvent exposed only upon changes in protein conformation. Phosphorylation and increased flux of nucleotide substrates served as two distinct modes by which EGF specified the cryptic cysteine residues that became solvent exposed and redox regulated. Because proteins that are structurally regulated by different RTKs or cellular perturbations are largely unique, these findings suggest that solvent exposure and redox regulation of cryptic cysteine residues contextually delineate redox signaling networks.

scholarly journals Receptor Tyrosine Kinases Fall into Distinct Classes Based on Their Inferred Signaling Networks

2013 ◽  
Vol 6 (284) ◽  
pp. ra58-ra58 ◽  
Author(s):  
J. P. Wagner ◽  
A. Wolf-Yadlin ◽  
M. Sevecka ◽  
J. K. Grenier ◽  
D. E. Root ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Signaling Networks

scholarly journals S-sulfhydration as a cellular redox regulation

2016 ◽  
Vol 36 (2) ◽  
Author(s):  
Małgorzata Iciek ◽  
Danuta Kowalczyk-Pachel ◽  
Anna Bilska-Wilkosz ◽  
Inga Kwiecień ◽  
Magdalena Górny ◽  
...  
Keyword(s):  
Protein Function ◽  
Redox Regulation ◽  
Biological Significance ◽  
Redox Signaling ◽  
Sulfur Species ◽  
Cellular Redox ◽  
Reactive Sulfur Species

This review is focused on formation and biological significance of hydropersulfides, i.e. S-sulfhydration process. Biogenesis and properties of reactive sulfur species and their role in redox signaling are presented. The effect of S-sulfhydration on protein function is discussed.

scholarly journals Downregulation of the Ras–Mitogen-Activated Protein Kinase Pathway by the EphB2 Receptor Tyrosine Kinase Is Required for Ephrin-Induced Neurite Retraction

2001 ◽  
Vol 21 (21) ◽  
pp. 7429-7441 ◽  
Author(s):  
Sabine Elowe ◽  
Sacha J. Holland ◽  
Sarang Kulkarni ◽  
Tony Pawson
Keyword(s):  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Mapk Pathway ◽  
Neuronal Cells ◽  
Mitogen Activated Protein Kinase ◽  
Neurite Retraction ◽  
Mitogen Activated Protein ◽  
Stimulation Of

ABSTRACT Activation of the EphB2 receptor tyrosine kinase by clustered ephrin-B1 induces growth cone collapse and neurite retraction in differentiated NG108 neuronal cells. We have investigated the cytoplasmic signaling events associated with EphB2-induced cytoskeletal reorganization in these neuronal cells. We find that unlike other receptor tyrosine kinases, EphB2 induces a pronounced downregulation of GTP-bound Ras and consequently of the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) pathway. A similar inhibition of the Ras-MAPK pathway was observed on stimulation of endogenous EphB2 in COS-1 cells. Inactivation of Ras, induced by ephrin B1 stimulation of NG108 neuronal cells, requires EphB2 tyrosine kinase activity and is blocked by a truncated form of p120-Ras GTPase-activating protein (p120-RasGAP), suggesting that EphB2 signals through the SH2 domain protein p120-RasGAP to inhibit the Ras-MAPK pathway. Suppression of Ras activity appears functionally important, since expression of a constitutively active variant of Ras impaired the ability of EphB2 to induce neurite retraction. In addition, EphB2 attenuated the elevation in ERK activation induced by attachment of NG108 cells to fibronectin, indicating that the EphB2 receptor can modulate integrin signaling to the Ras GTPase. These results suggest that a primary function of EphB2, a member of the most populous family of receptor tyrosine kinases, is to inactivate the Ras-MAPK pathway in a fashion that contributes to cytoskeletal reorganization and adhesion responses in neuronal growth cones.

scholarly journals Global profiling of distinct cysteine redox forms reveals wide-ranging redox regulation in C. elegans

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jin Meng ◽  
Ling Fu ◽  
Keke Liu ◽  
Caiping Tian ◽  
Ziyun Wu ◽  
...  
Keyword(s):  
Redox Regulation ◽  
Mapk Pathway ◽  
Model Organism ◽  
Redox Signaling ◽  
P38 Map Kinase ◽  
Redox Biology ◽  
C Elegans ◽  
Protein Functions ◽  
Redox Forms

AbstractPost-translational changes in the redox state of cysteine residues can rapidly and reversibly alter protein functions, thereby modulating biological processes. The nematode C. elegans is an ideal model organism for studying cysteine-mediated redox signaling at a network level. Here we present a comprehensive, quantitative, and site-specific profile of the intrinsic reactivity of the cysteinome in wild-type C. elegans. We also describe a global characterization of the C. elegans redoxome in which we measured changes in three major cysteine redox forms after H2O2 treatment. Our data revealed redox-sensitive events in translation, growth signaling, and stress response pathways, and identified redox-regulated cysteines that are important for signaling through the p38 MAP kinase (MAPK) pathway. Our in-depth proteomic dataset provides a molecular basis for understanding redox signaling in vivo, and will serve as a valuable and rich resource for the field of redox biology.

scholarly journals Akt-RSK-S6 Kinase Signaling Networks Activated by Oncogenic Receptor Tyrosine Kinases

2010 ◽  
Vol 3 (136) ◽  
pp. ra64-ra64 ◽  
Author(s):  
A. Moritz ◽  
Y. Li ◽  
A. Guo ◽  
J. Villen ◽  
Y. Wang ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Signaling Networks ◽  
Kinase Signaling ◽  
S6 Kinase
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