Crystal Structure of Hck in Complex with a Src Family–Selective Tyrosine Kinase Inhibitor

Schindler, Thomas; Sicheri, Frank; Pico, Alexander R.; Gazit, Aviv; Levitzki, Alexander; Kuriyan, John

doi:10.1016/s1097-2765(00)80357-3

Cited by 395 publications

(453 citation statements)

References 45 publications

Supporting

Mentioning

431

Contrasting

Order By: Relevance

“…3D), which could be explained by the likely induction of the closed conformation of SFKs upon the inhibition by LAT-Csk. This conformation stabilizes the activation loop in the inactive position where the activation loop tyrosine might be relatively difficult to access (48). PP2-inhibited Lyn is characterized by a very low level of Y507 phosphorylation (Fig.…”

Section: Inhibition Of Sfks Blocks Bcr Signaling Induced By a Bcr Ligandmentioning

confidence: 98%

Nonredundant Roles of Src-Family Kinases and Syk in the Initiation of B-Cell Antigen Receptor Signaling

Štěpánek

Dráber

Drobek

et al. 2013

The Journal of Immunology

View full text Add to dashboard Cite

When a BCR on a mature B cell is engaged by its ligand, the cell becomes activated, and the Ab-mediated immune response can be triggered. The initiation of BCR signaling is orchestrated by kinases of the Src and Syk families. However, the proximal BCR-induced phosphorylation remains incompletely understood. According to a model of sequential activation of kinases, Syk acts downstream of Src family kinases (SFKs). In addition, signaling independent of SFKs and initiated by Syk has been proposed. Both hypotheses lack sufficient evidence from relevant B cell models, mainly because of the redundancy of Src family members and the importance of BCR signaling for B cell development. We addressed this issue by analyzing controlled BCR triggering ex vivo on primary murine B cells and on murine and chicken B cell lines. Chemical and Csk-based genetic inhibitor treatments revealed that SFKs are required for signal initiation and Syk activation. In addition, ligand and anti-BCR Ab–induced signaling differ in their sensitivity to the inhibition of SFKs.

show abstract

Section: Inhibition Of Sfks Blocks Bcr Signaling Induced By a Bcr Ligandmentioning

confidence: 98%

Nonredundant Roles of Src-Family Kinases and Syk in the Initiation of B-Cell Antigen Receptor Signaling

Štěpánek

Dráber

Drobek

et al. 2013

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…For protein production, Sf9 cells (1 L) were grown in suspension in Grace's medium (Invitrogen) supplemented with 10% FBS and 50 μg/ml gentamycin. Sf9 cells were cultured to a density of 2 × 10 6 cells per mL and then co-infected with the Abl core and YopH baculoviruses at a multiplicity of infection (18). Upon purification, the protein was dialyzed against 20 mM Tris/ HCl, pH 8.3, containing 100 mM NaCl, and 3mM DTT.…”

Section: Dna Constructs and Protein Purificationmentioning

confidence: 99%

Abl N-Terminal Cap Stabilization of SH3 Domain Dynamics

et al. 2008

View full text Add to dashboard Cite

Crystal structures and other biochemical data indicate that the N-terminal cap (NCap) region of the Abelson tyrosine kinase (c-Abl) is important for maintaining the downregulated conformation of the kinase domain. The exact contributions that NCap makes in stabilizing the various intramolecular interactions within c-Abl are less clear. While the NCap appears important for locking the SH3/SH2 domains to the back of the kinase domain, there may be other more subtle elements of regulation. Hydrogen exchange (HX) and mass spectrometry (MS) were used to determine if the NCap contributes to intramolecular interactions involving the Abl SH3 domain. Under physiological conditions, the Abl SH3 domain underwent partial unfolding and its unfolding half-life was slowed during binding to the SH2-kinase linker, providing a unique assay to test NCap-induced stabilization of the SH3 domain in various constructs. The results showed that NCap stabilizes the dynamics of the SH3 domain in certain constructs but does not increase the relative affinity of the SH3 domain for the native SH2-kinase linker. The stabilization effect was absent in constructs of just NCap + SH3 but was obvious when the SH2 domain and the SH2-kinase linker were present. These results suggest that interactions between NCap and the SH3 domain can contribute to c-Abl stabilization in constructs that contain at least the SH2 domain, an effect that may partially compensate for the absence of the negative regulatory C-terminal tail found in the related Src family of kinases.The Abelson protein-tyrosine kinase (c-Abl) is a non-receptor tyrosine kinase ubiquitously expressed and highly conserved in metazoan evolution. c-Abl has many cellular functions including regulation of cell growth, survival, and differentiation, oxidative stress and DNAdamage responses, actin dynamics, and cell adhesion and migration (1,2). Fusion of the gene encoding c-Abl on chromosome 9 with the breakpoint cluster region (Bcr) gene on chromosome 22 results in the formation of a fusion protein, Bcr-Abl, which has constitutive protein-tyrosine kinase activity (3,4). The enhanced tyrosine kinase activity of the Bcr-Abl protein contributes to several disease states including chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL) (4). However regulation of both c-Abl and the BcrAbl fusion protein is still not well understood.The tyrosine kinase core of c-Abl is composed of a number of regions including an N-terminal cap (NCap) region, an SH3 domain, an SH2 domain, and a kinase domain (see Fig. 1A). Multiple intramolecular interactions involving these regions occur in c-Abl and these interactions are crucial for maintaining the downregulated state (5). Although the c-Abl core is very similar to Src-family tyrosine kinases (SFKs) in structure (1,6,7), significant differences † We are pleased to acknowledge generous financial support from the NIH: GM070590 (to J.R.E.) and CA101828 (to T.E.S.) *Address correspondence: John R. Engen 341 Mugar Life Sciences, The Barnett Instit...

show abstract

“…The crystal structures of inactive Hck complexed with the nucleoside analog PP1 (Protein Data Bank ID 1QCF, [10]), was used to generate the starting structures. For consistency, the sequence numbering from entry 1QCF was also adapted in the present work.…”

Section: Preparation Of Starting Structuresmentioning

confidence: 99%

“…The crystal structures of the inactive ("closed") forms of c-Src [6][7][8] and Hck [9,10] reveal that intramolecular interactions involving the SH2 and SH3 domains are essential for suppression of kinase activity despite being far away from the active site. In fact, these regulatory domains are bound to the distal side of the catalytic domain, restricting its conformational flexibility to hinder productive ATP binding.…”

Section: Introductionmentioning

confidence: 99%

“…Activation of Src-family kinases (SFKs) requires the disruption of the intramolecular interactions, the dephosphorylation of the C-terminal tail by phosphatases, or the binding of high-affinity ligands to the SH2 or the SH3 domain [9][10][11][12]. Additionally, the active conformation becomes stabilized upon autophosphorylation of Tyr416 (in Src) in the activation segment and is required for maximal kinase activity [13,14].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Effect of the SH3-SH2 domain linker sequence on the structure of Hck kinase

Meiselbach

Sticht

2010

J Mol Model

View full text Add to dashboard Cite

The coordination of activity in biological systems requires the existence of different signal transduction pathways that interact with one another and must be precisely regulated. The Src-family tyrosine kinases, which are found in many signaling pathways, differ in their physiological function despite their high overall structural similarity. In this context, the differences in the SH3-SH2 domain linkers might play a role for differential regulation, but the structural consequences of linker sequence are yet poorly understood. We have therefore performed comparative molecular dynamics simulations of wildtype Hck and of a mutant Hck, in which the SH3-SH2 domain linker is replaced by the sequence from the homologous kinase Lck. The simulations reveal that linker replacement does not only affect the orientation of the SH3 domain itself, but also leads to an alternative conformation of the activation segment in the Hck kinase domain. The sequence of the SH3-SH2 domain linker thus exerts a remote effect on the active site geometry and might therefore play a role in modulating the structure of the inactive kinase or for the fine-tuning of the activation process itself.

show abstract

Crystal Structure of Hck in Complex with a Src Family–Selective Tyrosine Kinase Inhibitor

Cited by 395 publications

References 45 publications

Nonredundant Roles of Src-Family Kinases and Syk in the Initiation of B-Cell Antigen Receptor Signaling

Nonredundant Roles of Src-Family Kinases and Syk in the Initiation of B-Cell Antigen Receptor Signaling

Abl N-Terminal Cap Stabilization of SH3 Domain Dynamics

Effect of the SH3-SH2 domain linker sequence on the structure of Hck kinase

Contact Info

Product

Resources

About