Two conserved cysteine residues are critical for the enzymic function of the human platelet-derived growth factor receptor-β: evidence for different roles of Cys-822 and Cys-940 in the kinase activity

Lee, Joon Won; Kim, Jee Eun; Park, Eun Jung; Kim, Jin Hyun; Lee, Chang-Hun; Lee, Seung Rock; Kwon, Jongbum

doi:10.1042/bj20040624

Cited by 13 publications

(10 citation statements)

References 33 publications

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“…A separate study examined the functional role of conserved cysteine residues located in the cytoplasmic domain of PDGFR. Treatment with the sulfhydryl-specific alkylating agent, N -ethylmaleimide (NEM), inhibits kinase activity and suggests the receptor contains critical cysteine residues located within its kinase domain (Lee et al, 2004). MS analysis identified two residues, Cys822 and Cys940, deemed necessary for receptor activity by in vitro assays using Ser mutants.…”

Section: Receptor Tyrosine Kinasesmentioning

confidence: 99%

Redox regulation of protein kinases

Truong

Carroll

2013

Critical Reviews in Biochemistry and Molecular Biology

151

105

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Protein kinases represent one of the largest families of genes found in eukaryotes. Kinases mediate distinct cellular processes ranging from proliferation, differentiation, survival, and apoptosis. Ligand-mediated activation of receptor kinases can lead to the production of endogenous H2O2 by membrane-bound NADPH oxidases. In turn, H2O2 can be utilized as a secondary messenger in signal transduction pathways. This review presents an overview of the molecular mechanisms involved in redox regulation of protein kinases and its effects on signaling cascades. In the first half, we will focus primarily on receptor tyrosine kinases (RTKs), whereas the latter will concentrate on downstream non-receptor kinases involved in relaying stimulant response. Select examples from the literature are used to highlight the functional role of H2O2 regarding kinase activity, as well as the components involved in H2O2 production and regulation during cellular signaling. In addition, studies demonstrating direct modulation of protein kinases by H2O2 through cysteine oxidation will be emphasized. Identification of these redox-sensitive residues may help uncover signaling mechanisms conserved within kinase subfamilies. In some cases, these residues can even be exploited as targets for the development of new therapeutics. Continued efforts in this field will further basic understanding of kinase redox regulation, and delineate the mechanisms involved in physiologic and pathological H2O2 responses.

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Section: Receptor Tyrosine Kinasesmentioning

confidence: 99%

Redox regulation of protein kinases

Truong

Carroll

2013

Critical Reviews in Biochemistry and Molecular Biology

151

105

View full text Add to dashboard Cite

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“…Protein spots were excised from silver-stained gels and subjected to trypsin digestion and MALDI-TOF analysis by essentially the same method as described previously [12] using the MS core facility of the Center for Cell Signaling Research, Ewha Woman's University.…”

Section: -D (Two-dimensional) Electrophoresis and Maldi-tof (Matrixamentioning

confidence: 99%

A proteomics approach for the identification of nucleophosmin and heterogeneous nuclear ribonucleoprotein C1/C2 as chromatin-binding proteins in response to DNA double-strand breaks

Lee

Park

Kim

et al. 2005

Biochemical Journal

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Double-strand breaks (DSBs) of chromosomal DNA trigger the cellular response that activates the pathways for DNA repair and cell-cycle checkpoints, and sometimes the pathways leading to cell death if the damage is too severe to be tolerated. Evidence indicates that, upon generation of DNA DSBs, many nuclear proteins that are involved in DNA repair and checkpoints are recruited to chromatin around the DNA lesions. In the present study we used a proteomics approach to identify DNA-damage-induced chromatin-binding proteins in a systematic way. Two-dimensional gel analysis for protein extracts of chromatin from DNA-damage-induced and control HeLa cells identified four proteins as the candidates for DNA-damage-induced chromatin-binding proteins. MALDI-TOF (matrix-assisted laser-desorption ionization-time-of-flight) MS analysis identified these proteins to be NPM (nucleophosmin), hnRNP (heterogeneous nuclear ribonucleoprotein) C1, hnRNP C2 and 37-kDa laminin-receptor precursor, and the identity of these proteins was further confirmed by immunoblot analysis with specific antibodies. We then demonstrated with chromatin-binding assays that NPM and hnRNP C1/C2, the abundant nuclear proteins with pleiotropic functions, indeed bind to chromatin in a DNA-damage-dependent manner, implicating these proteins in DNA repair and/or damage response. Immunofluorescence experiments showed that NPM, normally present in the nucleoli, is mobilized into the nucleoplasm after DNA damage, and that neither NPM nor hnRNP C1/C2 is actively recruited to the sites of DNA breaks. These results suggest that NPM and hnRNP C1/C2 may function at the levels of the global context of chromatin, rather than by specifically targeting the broken DNA.

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“…Herein, we have identified a redox-active highly conserved amino acid Mx (2) CWx (6) R motif located at the C-end of the kinase domain of Zap70 which is crucial for the regulation of Zap70 stability/activity. The oxidation of the cysteine within this motif appears to be crucial not only for Zap70, as its substitution in Src (C498A), Yes (C506A), Lyn (C479A), Lck (C475A), c-Ret (C376A), PDGFR-β (C940A), and FAK (C685A) affects kinase activity and in some cases also protein stability [ 24 , 30 , 31 , 34 – 39 ]. We have shown for the first time that in addition to the cysteine also the other conserved amino acids are crucial for protein stability.…”

Section: Resultsmentioning

confidence: 99%

A highly conserved redox-active Mx(2)CWx(6)R motif regulates Zap70 stability and activity

et al. 2017

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ζ-associated protein of 70 kDa (Zap70) is crucial for T-cell receptor (TCR) signaling. Loss of Zap70 in both humans and mice results in severe immunodeficiency. On the other hand, the expression of Zap70 in B-cell malignancies correlates with the severity of the disease. Because of its role in immune-related disorders, Zap70 has become a therapeutic target for the treatment of human diseases. It is well-established that the activity/expression of Zap70 is regulated by post-translational modifications of crucial amino acids including the phosphorylation of tyrosines and the ubiquitination of lysines. Here, we have investigated whether also oxidation of cysteine residues regulates Zap70 functions. We have identified C575 as a major sulfenylation site of Zap70. A C575A substitution results in protein instability, reduced activity, and increased dependency on the Hsp90/Cdc37 chaperone system. Indeed, Cdc37 overexpression reconstituted partially the expression but fully the function of Zap70C575A. C575 lies within a Mx(2)CWx(6)R motif which is highly conserved among almost all human tyrosine kinases. Mutation of any of the conserved amino acids, but not of a non-conserved residue preceding the cysteine, also results in Zap70 instability. Collectively, we have identified a new redox-active motif which is crucial for the regulation of Zap70 stability/activity. We believe that this motif has the potential to become a novel target for the development of therapeutic tools to modulate the expression/activity of kinases.

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Two conserved cysteine residues are critical for the enzymic function of the human platelet-derived growth factor receptor-β: evidence for different roles of Cys-822 and Cys-940 in the kinase activity

Cited by 13 publications

References 33 publications

Redox regulation of protein kinases

Redox regulation of protein kinases

A proteomics approach for the identification of nucleophosmin and heterogeneous nuclear ribonucleoprotein C1/C2 as chromatin-binding proteins in response to DNA double-strand breaks

A highly conserved redox-active Mx(2)CWx(6)R motif regulates Zap70 stability and activity

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