Substitution Studies of the Second Divalent Metal Cation Requirement of Protein Tyrosine Kinase CSK

Sun, Gongqin; Budde, Raymond J.A.

doi:10.1021/bi982793w

Cited by 52 publications

(60 citation statements)

References 34 publications

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“…There are also serine/threonine kinases that can be activated by Mn 2ϩ (37,40). This effect is most likely because of the binding of Mn 2ϩ instead of Mg 2ϩ to the second metalbinding site (31,39). At the same time, it was demonstrated that binding of Zn 2ϩ or Co 2ϩ at this second binding site is inhibitory in tyrosine kinases (39).…”

Section: Discussionmentioning

confidence: 99%

“…This effect is most likely because of the binding of Mn 2ϩ instead of Mg 2ϩ to the second metalbinding site (31,39). At the same time, it was demonstrated that binding of Zn 2ϩ or Co 2ϩ at this second binding site is inhibitory in tyrosine kinases (39). A striking similarity between the effects of divalent metal ions on ChaK1-cat activity and tyrosine kinases suggests that the catalytic mechanism of phosphotransfer in ␣-kinases and conventional protein kinases may be similar.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of the Protein Kinase Activity of TRPM7/ChaK1, a Protein Kinase Fused to the Transient Receptor Potential Ion Channel

Ryazanova¹,

Dorovkov²,

Ansari³

et al. 2004

Journal of Biological Chemistry

171

142

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Channel-kinase TRPM7/ChaK1 is a member of a recently discovered family of protein kinases called ␣-kinases that display no sequence homology to conventional protein kinases. It is an unusual bifunctional protein that contains an ␣-kinase domain fused to an ion channel. The TRPM7/ChaK1 channel has been characterized using electrophysiological techniques, and recent evidence suggests that it may play a key role in the regulation of magnesium homeostasis. However, little is known about its protein kinase activity. To characterize the kinase activity of TRPM7/ChaK1, we expressed the kinase catalytic domain in bacteria. ChaK1-cat is able to undergo autophosphorylation and to phosphorylate myelin basic protein and histone H3 on serine and threonine residues. The kinase is specific for ATP and cannot use GTP as a substrate. ChaK1-cat is insensitive to staurosporine (up to 0.1 mM) but can be inhibited by rottlerin. Because the kinase domain is physically linked to an ion channel, we investigated the effect of ions on ChaK1-cat activity. The kinase requires Mg 2؉ (optimum at 4 -10 mM) or Mn 2؉ (optimum at 3-5 mM), with activity in the presence of Mn 2؉ being 2 orders of magnitude higher than in the presence of Mg 2؉ . Zn 2؉ and Co 2؉ inhibited ChaK1-cat kinase activity. Ca 2؉ at concentrations up to 1 mM did not affect kinase activity. Considering intracellular ion concentrations, our results suggest that, among divalent metal ions, only Mg 2؉ can directly modulate TRPM7/ChaK1 kinase activity in vivo.A new family of protein kinases that do not display sequence homology to conventional eukaryotic protein kinases has been recently identified (1, 2). When mammalian and Caenorhabditis elegans elongation factor-2 kinases (eEF-2 1 kinases) were cloned, it was found that they do not display sequence homology to any conventional eukaryotic protein kinase (1). However, their catalytic domains appeared to be homologous to the catalytic domain of myosin heavy chain kinase A from Dictyostelium (3-5). Two more protein kinases with the same type of catalytic domain have been subsequently identified in Dictyostelium and have been called myosin heavy chain kinases B and C (6, 7). This new family of protein kinases was named ␣-kinase, because the existing evidence suggests that these protein kinases phosphorylate amino acids located within ␣-helices (2). This is different from conventional protein kinases that phosphorylate amino acids located within loops, turns, or regions with irregular structure (8). The ␣-kinase catalytic domain is characterized by several conserved motifs, which are different from the distinguishing sequence motifs that are found in conventional protein kinases (2). Surprisingly, the recently determined three-dimensional structure of the ␣-kinase catalytic domain revealed that, despite the lack of sequence homology, ␣-kinases have a fold that is very similar to conventional eukaryotic protein kinases (9).Five more human proteins with the ␣-kinase domain have been identified and cloned (2, 10, 11). Unexpectedly, it ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Characterization of the Protein Kinase Activity of TRPM7/ChaK1, a Protein Kinase Fused to the Transient Receptor Potential Ion Channel

Ryazanova¹,

Dorovkov²,

Ansari³

et al. 2004

Journal of Biological Chemistry

171

142

View full text Add to dashboard Cite

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“…Two cation binding sites in the active site of kinases have been defined, and binding of cations to the second, lower affinity site can either be stimulatory or inhibitory (25)(26)(27). It was previously reported that Her4 kinase activity is higher in the presence of Mn 2ϩ than with Mg 2ϩ , but that Mn 2ϩ concentrations Ͼ0.5 mM resulted in a reduction in Her4 kinase activity (28).…”

Section: Her4 Kinase Domain Is Activated By Dimerizing On the Surfacementioning

confidence: 99%

Her4 and Her2/neu Tyrosine Kinase Domains Dimerize and Activate in a Reconstituted in Vitro System

Monsey

Shen

Schlesinger

et al. 2010

Journal of Biological Chemistry

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Her4 (ErbB-4) and Her2/neu (ErbB-2) are receptor-tyrosine kinases belonging to the epidermal growth factor receptor (EGFR) family. Crystal structures of EGFR and Her4 kinase domains demonstrate kinase dimerization and activation through an allosteric mechanism. The kinase domains form an asymmetric dimer, where the C-lobe surface of one monomer contacts the N-lobe of the other monomer. EGFR kinase dimerization and activation in vitro was previously reported using a nickel-chelating lipid-liposome system, and we now apply this system to all other members of the EGFR family. Polyhistidinetagged Her4, Her2/neu, and Her3 kinase domains are bound to these nickel-liposomes and are brought to high local concentration, mimicking what happens to full-length receptors in vivo following ligand binding. Addition of nickel-liposomes to Her4 kinase domain results in 40-fold activation in kinase activity and marked enhancement of C-terminal tail autophosphorylation. Activation of Her4 shows a sigmoidal dependence on kinase concentration, consistent with a cooperative process requiring kinase dimerization. Her2/neu kinase activity is also activated by nickel-liposomes, and is increased further by heterodimerization with Her3 or Her4. The ability of Her3 and Her4 to heterodimerize and activate other family members is studied in vitro. Her3 kinase domain readily activates Her2/neu but is a poor activator of Her4, which differs from the prediction made by the asymmetric dimer model. Mutation of Her3 residues 952 ENI 954 to the corresponding sequence in Her4 enhanced the ability of Her3 to activate Her4, demonstrating that sequence differences on the C-lobe surface influence the heterodimerization and activation of ErbB kinase domains.Her4 (ErbB-4) and Her2/neu (ErbB-2) are members of the ErbB family of receptor-tyrosine kinases, with the founding member of this family being epidermal growth factor receptor (EGFR) 3 (1). The ErbB receptor-tyrosine kinases have an extracellular domain that is involved in ligand binding and receptor homo-or heterodimerization. Their intracellular domain consists of the juxtamembrane region, tyrosine kinase domain, and C-terminal tail. EGFR, Her2/neu, and Her4 contain catalytically active kinase domains, whereas Her3 (ErbB-3) has an inactive kinase domain and must heterodimerize with another ErbB receptor to signal (2). Several studies on ErbB heterodimerization have shown that Her2/neu is the preferred heterodimerization partner for the other three ErbB receptors (3-5). These studies also provide evidence for formation of EGFR-Her3 and EGFR-Her4 heterodimers in response to ligands that bind to Her3 or Her4 (3, 6). Both Her4 and Her2/neu play important roles in the development and the normal physiology of the cardiovascular and nervous systems (7,8). Her4 and Her2/neu knock-out mice both die around embryonic day 10.5 because of malformation of the cardiac trabeculae, and additionally, they are found to have abnormalities in the hindbrain or in sensory ganglia and motor nerves (9, 10). In human breast ...

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“…The group IIB metal ions are of great interest because Zn 21 , one of the most abundant divalent metal ions in living organisms, 1 is an essential cofactor in many metabolic enzymes and transcription factors. [2][3][4][5][6][7][8] On the other hand, Cd 21 and Hg 21 , which are in the same periodic group as Zn 21 , are known to be toxic to living organisms.…”

Section: Introductionmentioning

confidence: 99%

Force fields including charge transfer and local polarization effects: Application to proteins containing multi/heavy metal ions

Sakharov

Lim

2008

J Comput Chem

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The question whether molecular dynamics (MD) simulations can yield reliable structural and dynamical properties of metalloproteins depend on the accuracy of the force field, i.e., the potential energy function (PEF) and associated parameters modeling the interactions of the metal ion of interest with water and protein ligands. Previously, we had developed a CTPOL PEF for protein simulations of Zn(2+) bound to Cys(-) and/or His(0) that includes charge transfer and local polarization effects as well as metal van der Waals parameters that reproduce the structural and thermodynamical properties of 22 dications. Here, we evaluate if the CTPOL PEF and the new metal parameters (referred to as the CTPOLa force field) can be applied to proteins containing polynuclear metal-binding sites and heavy toxic metal ions, using the CdZn(2)-Cys(9) beta-domain of rat liver metallothionein-2 and the Hg(2+)-bound 18-residue peptide from MerP as test systems. Using the CTPOLa force field, simulations of the beta-domain of rat liver metallothionein-2 totaling 19 ns could preserve the experimentally observed CdZn(2)-Cys(9) complex geometry and overall protein structure, whereas simulations neglecting charge transfer and local polarization effects could not. However, the CTPOLa force field cannot reproduce the experimentally observed linear bicoordination of Hg(2+) in the MerP peptide without adding an angular restraint to the CTPOL PEF to correct the angle distribution about Hg(2+). Thus, the force fields presented herein for the group IIB metal ions can be applied to simulation studies of proteins containing polynuclear metal-binding sites and heavy metal ions in aqueous solution. PEF neglecting charge transfer and local polarization effects in conjunction with vdW parameters adjusted to reproduce the structural and thermodynamical properties of only the metal ion in question could not yield an accurate representation of the metal-binding site and overall protein structure.

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Substitution Studies of the Second Divalent Metal Cation Requirement of Protein Tyrosine Kinase CSK

Abstract: In addition to a magnesium ion needed to form the ATP-Mg complex, we have previously determined that at least one more free Mg 2+ ion is essential for the activation of the protein tyrosine kinase, Csk [

Cited by 52 publications

References 34 publications

Characterization of the Protein Kinase Activity of TRPM7/ChaK1, a Protein Kinase Fused to the Transient Receptor Potential Ion Channel

Characterization of the Protein Kinase Activity of TRPM7/ChaK1, a Protein Kinase Fused to the Transient Receptor Potential Ion Channel

Her4 and Her2/neu Tyrosine Kinase Domains Dimerize and Activate in a Reconstituted in Vitro System

Force fields including charge transfer and local polarization effects: Application to proteins containing multi/heavy metal ions

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