The Protein Kinase C Inhibitor Bisindolyl Maleimide 2 Binds with Reversed Orientations to Different Conformations of Protein Kinase A

Gaßel, Michael; Breitenlechner, C.B.; König, Norbert; Huber, Robert; Engh, Richard A.; Bossemeyer, Dirk

doi:10.1074/jbc.m314082200

Cited by 40 publications

(48 citation statements)

References 46 publications

(41 reference statements)

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“…To elucidate the signaling mechanisms by which intracellular M. leprae induce MEK-independent Erk1͞2 activation, 30-dayinfected Schwann cells were serum-starved for 48 h with known inhibitors of major signaling pathways. We found that PKC paninhibitor (PKCI) bisindolyl maleimide-I (20 nM) (26), in the presence of MEKI and PI3KI, significantly blocked M. lepraeinduced phosphorylation of Erk1͞2 (Fig. 4A), suggesting a critical role of PKC in MEK-independent activation of Erk1͞2.…”

Section: Resultsmentioning

confidence: 94%

Insights into regulation of human Schwann cell proliferation by Erk1/2 via a MEK-independent and p56Lck-dependent pathway from leprosy bacilli

Tapinos

Rambukkana

2005

Proc. Natl. Acad. Sci. U.S.A.

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Activation of extracellular signal-regulated kinase (Erk) 1͞2, which plays a critical role in diverse cellular processes, including cell proliferation, is known to be mediated by the canonical Rafmitogen-activated protein kinase kinase (MEK) kinase cascade. Alternative MEK-independent signaling pathways for Erk1͞2 activation in mammalian cells are not known. During our studies of human primary Schwann cell response to long-term infection of Mycobacterium leprae, the causative organism of leprosy, we identified that intracellular M. leprae activated Erk1͞2 directly by lymphoid cell kinase (p56Lck), a Src family member, by means of a PKC-dependent and MEK-independent signaling pathway. Activation of this signaling induced nuclear accumulation of cyclin D1, G 1͞S-phase progression, and continuous proliferation, but without transformation. Thus, our data reveal a previously unknown signaling mechanism of glial cell proliferation, which might play a role in dedifferentiation as well as nerve regeneration and degeneration. Our findings may also provide a potential mechanism by which an obligate intracellular bacterial pathogen like M. leprae subverts nervous system signaling to propagate its cellular niche for colonization and long-term bacterial survival.cell signaling ͉ cell cycle ͉ peripheral nerve ͉ human ͉ Mycobacterium leprae

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Section: Resultsmentioning

confidence: 94%

Insights into regulation of human Schwann cell proliferation by Erk1/2 via a MEK-independent and p56Lck-dependent pathway from leprosy bacilli

Tapinos

Rambukkana

2005

Proc. Natl. Acad. Sci. U.S.A.

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“…selectivity. 43 The residues that make up the binding site region in each of the kinases under consideration are indicated in Figure 2.…”

Section: Resultsmentioning

confidence: 99%

Can MM‐PBSA calculations predict the specificities of protein kinase inhibitors?

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Bates

2006

J Comput Chem

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An application of the molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) protocol to the prediction of protein kinase inhibitor selectivity is presented. Six different inhibitors are placed in equivalent orientations in each of six different receptors. Fully solvated molecular dynamics is then run for 1 ns on each of the 36 complexes, and the resulting trajectories scored, using the implicit solvent model. The results show some correlation with experimentally-determined specificities; anomalies may be attributed to a variety of causes, including difficulties in quantifying induced fit penalties and variabilities in normal modes calculations. Decomposing interaction energies on a per-residue basis yields more useful insights into the natures of the binding modes and suggests that the real value of such calculations lies in understanding interactions rather than outright prediction.

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“…This phenomenon is visible in two of the PKAR5 complexes, and the question arises whether the peptide flip could be related to a specific mutation of PKAR5. Notably, we observed this phenomenon before in the PKA staurosporine complex (1STC) and in one of the two molecules in the asymmetric unit of a PKAB3 Bim2 complex (35).…”

Section: Effect Of the Mutations On Y-27632mentioning

confidence: 98%

Structural Analysis of Protein Kinase A Mutants with Rho-kinase Inhibitor Specificity

Bonn

Herrero

Breitenlechner

et al. 2006

Journal of Biological Chemistry

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Controlling aberrant kinase-mediated cellular signaling is a major strategy in cancer therapy; successful protein kinase inhibitors such as Tarceva and Gleevec verify this approach. Specificity of inhibitors for the targeted kinase(s), however, is a crucial factor for therapeutic success. Based on homology modeling, we previously identified four amino acids in the active site of Rho-kinase that likely determine inhibitor specificities observed for Rho-kinase relative to protein kinase A (PKA) (in PKA numbering: T183A, L49I, V123M, and E127D), and a fifth (Q181K) that played a surprising role in PKA-PKB hybrid proteins. We have systematically mutated these residues in PKA to their counterparts in Rho-kinase, individually and in combination. Using four Rho-kinase-specific, one PKA-specific, and one pan-kinase-specific inhibitor, we measured the inhibitor-binding properties of the mutated proteins and identify the roles of individual residues as specificity determinants. Two combined mutant proteins, containing the combination of mutations T183A and L49I, closely mimic Rho-kinase. Kinetic results corroborate the hypothesis that side-chain identities form the major determinants of selectivity. An unexpected result of the analysis is the consistent contribution of the individual mutations by simple factors. Crystal structures of the surrogate kinase inhibitor complexes provide a detailed basis for an understanding of these selectivity determinant residues. The ability to obtain kinetic and structural data from these PKA mutants, combined with their Rho-kinase-like selectivity profiles, make them valuable for use as surrogate kinases for structure-based inhibitor design.Phosphorylation via protein kinases is responsible for a large part of cellular signal transduction and is described as a universal regulatory mechanism (1, 2). Perturbation of kinase-mediated signaling pathways results in a number of diseases, including diabetes, cancer, and inflammation (3, 4). Because most protein kinases reside in the cell in an inactive state and are activated by signal transduction processes, many diseases are triggered by overactivation of protein kinases via mutation, overexpression, or malfunctioning cellular inhibition.The human genome encodes some 518 protein kinases (5) that are notably different in how their catalysis is regulated but share a catalytic domain conserved in sequence and structure (6, 7). The latter consists of 250 -300 amino acids, binds substrate and cosubstrate, and catalyzes the phosphorylation reaction.This catalytic domain, together with less conserved surrounding sites, has been the focus of inhibitor design that has exploited differences in kinase structure and pliability to achieve selectivity. Many drugs that target protein kinases are in clinical trials, and some have already been approved, such as the Abl kinase inhibitor Gleevec for therapy against chronic myelogenous leukemia (8) and Tarceva (erlotinib) against nonsmall cell lung cancer (9). The first protein kinase inhibitor that passed the cli...

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The Protein Kinase C Inhibitor Bisindolyl Maleimide 2 Binds with Reversed Orientations to Different Conformations of Protein Kinase A

Cited by 40 publications

References 46 publications

Insights into regulation of human Schwann cell proliferation by Erk1/2 via a MEK-independent and p56Lck-dependent pathway from leprosy bacilli

Insights into regulation of human Schwann cell proliferation by Erk1/2 via a MEK-independent and p56Lck-dependent pathway from leprosy bacilli

Can MM‐PBSA calculations predict the specificities of protein kinase inhibitors?

Structural Analysis of Protein Kinase A Mutants with Rho-kinase Inhibitor Specificity

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