scholarly journals Dynamic Equilibrium of the Aurora A Kinase Activation Loop Revealed by Single-Molecule Spectroscopy

2017 ◽  
Vol 56 (38) ◽  
pp. 11409-11414 ◽  
Author(s):  
James A. H. Gilburt ◽  
Hajrah Sarkar ◽  
Peter Sheldrake ◽  
Julian Blagg ◽  
Liming Ying ◽  
...  
Keyword(s):  
Single Molecule ◽  
Dynamic Equilibrium ◽  
Activation Loop ◽  
Single Molecule Spectroscopy ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Kinase Activation

scholarly journals Dynamic Equilibrium of the Aurora A Kinase Activation Loop Revealed by Single-Molecule Spectroscopy

2017 ◽  
Vol 129 (38) ◽  
pp. 11567-11572
Author(s):  
James A. H. Gilburt ◽  
Hajrah Sarkar ◽  
Peter Sheldrake ◽  
Julian Blagg ◽  
Liming Ying ◽  
...  
Keyword(s):  
Single Molecule ◽  
Dynamic Equilibrium ◽  
Activation Loop ◽  
Single Molecule Spectroscopy ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Kinase Activation

scholarly journals Detection of Ligand‐induced Conformational Changes in the Activation Loop of Aurora‐A Kinase by PELDOR Spectroscopy

ChemistryOpen ◽  
2016 ◽  
Vol 5 (6) ◽  
pp. 531-534 ◽  
Author(s):  
Selena G. Burgess ◽  
Maria Grazia Concilio ◽  
Richard Bayliss ◽  
Alistair J. Fielding
Keyword(s):  
Conformational Changes ◽  
Activation Loop ◽  
Aurora A Kinase ◽  
Aurora A

scholarly journals Insights into Aurora-A Kinase Activation Using Unnatural Amino Acids Incorporated by Chemical Modification

2013 ◽  
Vol 8 (10) ◽  
pp. 2184-2191 ◽  
Author(s):  
Fiona C. Rowan ◽  
Meirion Richards ◽  
Rachel A. Bibby ◽  
Andrew Thompson ◽  
Richard Bayliss ◽  
...  
Keyword(s):  
Amino Acids ◽  
Chemical Modification ◽  
Unnatural Amino Acids ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Kinase Activation

scholarly journals Aurora A kinase activation: Different means to different ends

2021 ◽  
Vol 220 (9) ◽  
Author(s):  
Nicolas Tavernier ◽  
Frank Sicheri ◽  
Lionel Pintard
Keyword(s):  
Spindle Assembly ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Serine Threonine Kinase ◽  
Kinase Activation ◽  
Mitotic Entry ◽  
Threonine Kinase ◽  
Molecular Studies ◽  
Signaling Events ◽  
Cellular Context

Aurora A is a serine/threonine kinase essential for mitotic entry and spindle assembly. Recent molecular studies have revealed the existence of multiple, distinct mechanisms of Aurora A activation, each occurring at specific subcellular locations, optimized for cellular context, and primed by signaling events including phosphorylation and oxidation.

scholarly journals A Focused Small-Molecule Screen Identifies PHA-680626 as an Amphosteric Inhibitor Disrupting the Interaction between Aurora-A and N-Myc

2021 ◽  
Author(s):  
Fani Souvalidou ◽  
Dalila Boi ◽  
Roberta Montanari ◽  
Federica Polverino ◽  
Grazia Marini ◽  
...  
Keyword(s):  
Cell Lines ◽  
Small Molecule ◽  
Conformational Changes ◽  
Neuroblastoma Cells ◽  
Activation Loop ◽  
Aurora A ◽  
Kinase Activation ◽  
Mitotic Kinase ◽  
Gene Coding

Neuroblastoma is a severe childhood disease, accounting for ≈10% of all infant cancers. The amplification of the MYCN gene, coding for the N-Myc transcriptional factor, is an essential marker correlated with tumor progression and poor prognosis. In neuroblastoma cells, the mitotic kinase Aurora-A (AURKA), also frequently overexpressed in cancer, prevents N-Myc degradation, by directly binding to a highly conserved N-Myc region, i.e. Myc Box I. As a result, elevated levels of N-Myc, which are required for the growth of MYCN amplified cells, are observed. During the last years, it has been demonstrated that the ATP competitive inhibitors of AURKA CD532, MLN8054 and Alisertib also cause essential conformational changes in the structure of the activation loop of the kinase that prevent N-Myc binding, thus impairing the formation of the AURKA/N-Myc complex. In this study, starting from a screening of crystal structures of AURKA in complex with known inhibitors, we identified additional compounds affecting the conformation of the kinase activation loop. We assessed the ability of such compounds to disrupt the interaction between AURKA and N-Myc in vitro, using Surface Plasmon Resonance competition assays, and in tumor cell lines overexpressing MYCN, by performing Proximity Ligation Assays. Finally, their effects on N-Myc cellular levels and cell viability were investigated. Our results, identifying PHA-680626 as an amphosteric inhibitor both in vitro and MYCN overexpressing cell lines, expand the repertoire of known conformational disrupting inhibitors of the AURKA/N-Myc complex, and confirm that altering the conformation of the activation loop of AURKA with a small molecule is an effective strategy to destabilize the AURKA/N-Myc interaction in neuroblastoma cancer cells.

scholarly journals Quantitative conformational profiling of kinase inhibitors reveals origins of selectivity for Aurora kinase activation states

2018 ◽  
Vol 115 (51) ◽  
pp. E11894-E11903 ◽  
Author(s):  
Eric W. Lake ◽  
Joseph M. Muretta ◽  
Andrew R. Thompson ◽  
Damien M. Rasmussen ◽  
Abir Majumdar ◽  
...  
Keyword(s):  
Large Scale ◽  
Kinase Inhibitors ◽  
Conformational Dynamics ◽  
Aurora Kinase ◽  
Activation Loop ◽  
Aurora A ◽  
Kinase Activation ◽  
Mitotic Kinase ◽  
Conformational Preferences ◽  
Wide Range

Protein kinases undergo large-scale structural changes that tightly regulate function and control recognition by small-molecule inhibitors. Methods for quantifying the conformational effects of inhibitors and linking them to an understanding of selectivity patterns have long been elusive. We have developed an ultrafast time-resolved fluorescence methodology that tracks structural movements of the kinase activation loop in solution with angstrom-level precision, and can resolve multiple structural states and quantify conformational shifts between states. Profiling a panel of clinically relevant Aurora kinase inhibitors against the mitotic kinase Aurora A revealed a wide range of conformational preferences, with all inhibitors promoting either the active DFG-in state or the inactive DFG-out state, but to widely differing extents. Remarkably, these conformational preferences explain broad patterns of inhibitor selectivity across different activation states of Aurora A, with DFG-out inhibitors preferentially binding Aurora A activated by phosphorylation on the activation loop, which dynamically samples the DFG-out state, and DFG-in inhibitors binding preferentially to Aurora A constrained in the DFG-in state by its allosteric activator Tpx2. The results suggest that many inhibitors currently in clinical development may be capable of differentiating between Aurora A signaling pathways implicated in normal mitotic control and in melanoma, neuroblastoma, and prostate cancer. The technology is applicable to a wide range of clinically important kinases and could provide a wealth of valuable structure–activity information for the development of inhibitors that exploit differences in conformational dynamics to achieve enhanced selectivity.

scholarly journals Arpc1b, a centrosomal protein, is both an activator and substrate of Aurora A

2010 ◽  
Vol 190 (1) ◽  
pp. 101-114 ◽  
Author(s):  
Poonam R. Molli ◽  
Da-Qiang Li ◽  
Rozita Bagheri-Yarmand ◽  
Suresh B. Pakala ◽  
Hiroshi Katayama ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Centrosome Amplification ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Independent Manner ◽  
Centrosomal Protein ◽  
Kinase Activation

Here we provide evidence in support of an inherent role for Arpc1b, a component of the Arp2/3 complex, in regulation of mitosis and demonstrate that its depletion inhibits Aurora A activation at the centrosome and impairs the ability of mammalian cells to enter mitosis. We discovered that Arpc1b colocalizes with γ-tubulin at centrosomes and stimulates Aurora A activity. Aurora A phosphorylates Arpc1b on threonine 21, and expression of Arpc1b but not a nonphosphorylatable Arpc1b mutant in mammalian cells leads to Aurora A kinase activation and abnormal centrosome amplification in a Pak1-independent manner. Together, these findings reveal a new function for Arpc1b in centrosomal homeostasis. Arpc1b is both a physiological activator and substrate of Aurora A kinase and these interactions help to maintain mitotic integrity in mammalian cells.

scholarly journals PHA-680626 Is an Effective Inhibitor of the Interaction between Aurora-A and N-Myc

2021 ◽  
Vol 22 (23) ◽  
pp. 13122
Author(s):  
Dalila Boi ◽  
Fani Souvalidou ◽  
Davide Capelli ◽  
Federica Polverino ◽  
Grazia Marini ◽  
...  
Keyword(s):  
Cell Lines ◽  
Conformational Changes ◽  
Neuroblastoma Cells ◽  
Activation Loop ◽  
Aurora A ◽  
Kinase Activation ◽  
Proximity Ligation ◽  
Mitotic Kinase ◽  
Gene Coding

Neuroblastoma is a severe childhood disease, accounting for ~10% of all infant cancers. The amplification of the MYCN gene, coding for the N-Myc transcription factor, is an essential marker correlated with tumor progression and poor prognosis. In neuroblastoma cells, the mitotic kinase Aurora-A (AURKA), also frequently overexpressed in cancer, prevents N-Myc degradation by directly binding to a highly conserved N-Myc region. As a result, elevated levels of N-Myc are observed. During recent years, it has been demonstrated that some ATP competitive inhibitors of AURKA also cause essential conformational changes in the structure of the activation loop of the kinase that prevents N-Myc binding, thus impairing the formation of the AURKA/N-Myc complex. In this study, starting from a screening of crystal structures of AURKA in complexes with known inhibitors, we identified additional compounds affecting the conformation of the kinase activation loop. We assessed the ability of such compounds to disrupt the interaction between AURKA and N-Myc in vitro, using Surface Plasmon Resonance competition assays, and in tumor cell lines overexpressing MYCN, by performing Proximity Ligation Assays. Finally, their effects on N-Myc cellular levels and cell viability were investigated. Our results identify PHA-680626 as an amphosteric inhibitor both in vitro and in MYCN overexpressing cell lines, thus expanding the repertoire of known conformational disrupting inhibitors of the AURKA/N-Myc complex and confirming that altering the conformation of the activation loop of AURKA with a small molecule is an effective strategy to destabilize the AURKA/N-Myc interaction in neuroblastoma cancer cells.

Ancient origins of allosteric activation in a Ser-Thr kinase

Science ◽  
2020 ◽  
Vol 367 (6480) ◽  
pp. 912-917 ◽  
Author(s):  
Adelajda Hadzipasic ◽  
Christopher Wilson ◽  
Vy Nguyen ◽  
Nadja Kern ◽  
Chansik Kim ◽  
...  
Keyword(s):  
Activation Mechanism ◽  
Activation Loop ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Allosteric Activation ◽  
Fundamental Interest ◽  
Evolutionary Path ◽  
Cellular Events ◽  
Sequence Reconstruction ◽  
Allosteric Activator

A myriad of cellular events are regulated by allostery; therefore, evolution of this process is of fundamental interest. Here, we use ancestral sequence reconstruction to resurrect ancestors of two colocalizing proteins, Aurora A kinase and its allosteric activator TPX2 (targeting protein for Xklp2), to experimentally characterize the evolutionary path of allosteric activation. Autophosphorylation of the activation loop is the most ancient activation mechanism; it is fully developed in the oldest kinase ancestor and has remained stable over 1 billion years of evolution. As the microtubule-associated protein TPX2 appeared, efficient kinase binding to TPX2 evolved, likely owing to increased fitness by virtue of colocalization. Subsequently, TPX2-mediated allosteric kinase regulation gradually evolved. Surprisingly, evolution of this regulation is encoded in the kinase and did not arise by a dominating mechanism of coevolution.

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