Protein-tyrosine kinase CAKβ/PYK2 is activated by binding Ca2+/calmodulin to FERM F2 α2 helix and thus forming its dimer

2008 ◽  
Vol 410 (3) ◽  
pp. 513-523 ◽  
Author(s):  
Takayuki Kohno ◽  
Eiko Matsuda ◽  
Hiroko Sasaki ◽  
Terukatsu Sasaki
Keyword(s):  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Kinase Domain ◽  
Ferm Domain ◽  
Protein Tyrosine ◽  
Calmodulin Binding ◽  
Tyrosine Kinase 2 ◽  
Mutant Sequence ◽  
A Cell ◽  
Α Helix

CAKβ (cell adhesion kinase β)/PYK2 (proline-rich tyrosine kinase 2) is the second protein-tyrosine kinase of the FAK (focal adhesion kinase) subfamily. It is different from FAK in that it is activated following an increase in cytoplasmic free Ca2+. In the present study we have investigated how Ca2+ activates CAKβ/PYK2. Calmodulin-agarose bound CAKβ/PYK2, but not FAK, in the presence of CaCl2. An α-helix (F2-α2) present in the FERM (band four-point-one, ezrin, radixin, moesin homology) F2 subdomain of CAKβ/PYK2 was the binding site of Ca2+/calmodulin; a mutant of this region, L176A/Q177A (LQ/AA) CAKβ/PYK2, bound to Ca2+/calmodulin much less than the wild-type. CAKβ/PYK2 is known to be prominently tyrosine phosphorylated when overexpressed from cDNA. The enhanced tyrosine phosphorylation was inhibited by W7, an inhibitor of calmodulin, and by a cell-permeable Ca2+ chelator and was almost defective in the LQ/AA-mutant CAKβ/PYK2. CAKβ/PYK2 formed a homodimer on binding of Ca2+/calmodulin, which might then induce a conformational change of the kinase, resulting in transphosphorylation within the dimer. The dimer was formed at a free-Ca2+ concentration of 8–12 μM and was stable at 500 nM Ca2+, but dissociated to a monomer in a Ca2+-free buffer. The dimer formation of CAKβ/PYK2 FERM domain was partially defective in the LQ/AA-mutant FERM domain and was blocked by W7 and by a synthetic peptide with amino acids 168–188 of CAKβ/PYK2, but not by a peptide with its LQ/AA-mutant sequence. It is known that the F2-α2 helix is found immediately adjacent to a hydrophobic pocket in the FERM F2 lobe, which locks, in the autoinhibited FAK, the C-lobe of the kinase domain. Our results indicate that Ca2+/calmodulin binding to the FERM F2-α2 helix of CAKβ/PYK2 releases its kinase domain from autoinhibition by forming a dimer.

Functional dissection of p56lck, a protein tyrosine kinase which mediates interleukin-2-induced activation of the c-fos gene

1994 ◽  
Vol 14 (9) ◽  
pp. 5812-5819
Author(s):  
H Shibuya ◽  
K Kohu ◽  
K Yamada ◽  
E L Barsoumian ◽  
R M Perlmutter ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Gene Activation ◽  
Interleukin 2 ◽  
Kinase Domain ◽  
Tyrosine Kinase Domain ◽  
Amino Acid Residues ◽  
Protein Tyrosine ◽  
Responsive Element

Members of the newly identified receptor family for cytokines characteristically lack the intrinsic protein tyrosine kinase domain that is a hallmark of other growth factor receptors. Instead, accumulating evidence suggests that these receptors utilize nonreceptor-type protein tyrosine kinases for downstream signal transduction by cytokines. We have shown previously that the interleukin-2 receptor beta-chain interacts both physically and functionally with a Src family member, p56lck, and that p56lck activation leads to induction of the c-fos gene. However, the mechanism linking p56lck activation with c-fos induction remains unelucidated. In the present study, we systematically examined the extent of c-fos promoter activation by expression of a series of p56lck mutants, using a transient cotransfection assay. The results define a set of the essential amino acid residues that regulate p56lck induction of the c-fos promoter. We also provide evidence that the serum-responsive element and sis-inducible element are both targets through which p56lck controls c-fos gene activation.

scholarly journals Inhibition of Osteoclast Function by Adenovirus Expressing Antisense Protein-tyrosine Kinase 2

2000 ◽  
Vol 276 (10) ◽  
pp. 7484-7492 ◽  
Author(s):  
Le T. Duong ◽  
Ichiro Nakamura ◽  
Päivi T. Lakkakorpi ◽  
Lorraine Lipfert ◽  
Andrew J. Bett ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Protein Tyrosine ◽  
Tyrosine Kinase 2 ◽  
Osteoclast Function ◽  
Protein Tyrosine Kinase 2

The amino-terminal 14 amino acids of v-src can functionally replace the extracellular and transmembrane domains of v-erbB

1991 ◽  
Vol 11 (9) ◽  
pp. 4760-4770
Author(s):  
M McMahon ◽  
R C Schatzman ◽  
J M Bishop
Keyword(s):  
Membrane Protein ◽  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Transmembrane Domain ◽  
Transmembrane Protein ◽  
Integral Membrane Protein ◽  
Kinase Domain ◽  
Protein Tyrosine ◽  
Amino Terminal ◽  
Transforming Activity

The retroviral oncogene v-erbB encodes a truncated form of the receptor for epidermal growth factor, an integral membrane protein-tyrosine kinase. By contrast, the oncogene v-src encodes a protein-tyrosine kinase that is a peripheral membrane protein. The morphologies and spectra of cells transformed by these two oncogenes differ. In an effort to identify the functional determinant(s) of these differences, we constructed and tested first deletion mutants of v-erbB and then chimeras between v-src and v-erbB. As reported previously, the absence of any membrane anchorage eliminated transformation by v-erbB. Anchorage of the cytoplasmic kinase domain of v-erbB to membranes with amino-terminal portions of the v-src protein permitted transformation. The phenotype and spectrum of transformation were those expected for v-erbB rather than for v-src. The transforming chimeras lost their biological activity if the signal for myristylation at the amino terminus of v-src was compromised by mutation. Biochemical fractionations revealed a correlation between transforming activity and the association of chimeric gene products with the membrane fraction of the cell. For reasons not yet apparent, the combined presence of membrane anchorage domains of v-src, and the transmembrane domain of v-erbB in the same chimera typically (but not inevitably) impeded transformation. Our results suggest that the specificity of transformation by v-erbB resides in the selection of substrates by the cytoplasmic domain of the gene product. The protein retains access to those substrates even when anchored to the membrane in the manner of a peripheral rather than a transmembrane protein.

scholarly journals ZAP-70 Protein Tyrosine Kinase Is Constitutively Targeted to the T Cell Cortex Independently of its SH2 Domains

1997 ◽  
Vol 137 (7) ◽  
pp. 1639-1649 ◽  
Author(s):  
Russell D.J. Huby ◽  
Makio Iwashima ◽  
Arthur Weiss ◽  
Steven C. Ley
Keyword(s):  
T Cell ◽  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Kinase Domain ◽  
Cell Cortex ◽  
Protein Tyrosine ◽  
Sh2 Domains ◽  
Anchoring Protein ◽  
High Concentrations ◽  
Lck Protein

ZAP-70 is a nonreceptor protein tyrosine kinase that is essential for signaling via the T cell antigen receptor (TCR). ZAP-70 becomes phosphorylated and activated by LCK protein tyrosine kinase after interaction of its two NH2-terminal SH2 domains with tyrosine-phosphorylated subunits of the activated TCR. In this study, the localization of ZAP-70 was investigated by immunofluorescence and confocal microscopy. ZAP-70 was found to be localized to the cell cortex in a diffuse band under the plasma membrane in unstimulated T cells, and this localization was not detectably altered by TCR stimulation. Analysis of mutants indicated that ZAP-70 targeting was independent of its SH2 domains but required its active kinase domain. The specific compartmentalization of ZAP-70 suggests that it may interact with an anchoring protein in the cell cortex via its hinge or kinase domains. It is likely that the maintenance of high concentrations of ZAP-70 at the cell cortex, that only has to move a short distance to interact with phophorylated TCR subunits, facilitates rapid initiation of signaling by the TCR. In addition, as the major increase in tyrosine phosphorylation induced by the TCR also occurs at the cell cortex (Ley, S.C., M. Marsh, C.R. Bebbington, K. Proudfoot, and P. Jordan. 1994. J. Cell. Biol. 125:639–649), ZAP-70 may be localized close to its downstream targets.

scholarly journals Crystal structure of the EphA4 protein tyrosine kinase domain in the apo- and dasatinib-bound state

FEBS Letters ◽  
2011 ◽  
Vol 585 (22) ◽  
pp. 3593-3599 ◽  
Author(s):  
Carine Farenc ◽  
Patrick H.N. Celie ◽  
Cornelis P. Tensen ◽  
Iwan J.P. de Esch ◽  
Gregg Siegal
Keyword(s):  
Crystal Structure ◽  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Bound State ◽  
Kinase Domain ◽  
Tyrosine Kinase Domain ◽  
Protein Tyrosine

Structure of the protein tyrosine kinase domain of C-terminal Src kinase (CSK) in complex with staurosporine 1 1Edited by I. A. Wilson

1999 ◽  
Vol 285 (2) ◽  
pp. 713-725 ◽  
Author(s):  
Marieke B.A.C Lamers ◽  
Alfred A Antson ◽  
Roderick E Hubbard ◽  
Richard K Scott ◽  
David H Williams
Keyword(s):  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Src Kinase ◽  
Kinase Domain ◽  
Tyrosine Kinase Domain ◽  
Protein Tyrosine
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