scholarly journals Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Adelajda Zorba ◽  
Vanessa Buosi ◽  
Steffen Kutter ◽  
Nadja Kern ◽  
Francesco Pontiggia ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Regulatory Mechanism ◽  
Aurora A Kinase ◽  
Active Conformation ◽  
Aurora A ◽  
Allosteric Activation ◽  
X Ray ◽  
X Ray Crystallography ◽  
Functional Assays ◽  
Spindle Microtubules

We elucidate the molecular mechanisms of two distinct activation strategies (autophosphorylation and TPX2-mediated activation) in human Aurora A kinase. Classic allosteric activation is in play where either activation loop phosphorylation or TPX2 binding to a conserved hydrophobic groove shifts the equilibrium far towards the active conformation. We resolve the controversy about the mechanism of autophosphorylation by demonstrating intermolecular autophosphorylation in a long-lived dimer by combining X-ray crystallography with functional assays. We then address the allosteric activation by TPX2 through activity assays and the crystal structure of a domain-swapped dimer of dephosphorylated Aurora A and TPX21−25. While autophosphorylation is the key regulatory mechanism in the centrosomes in the early stages of mitosis, allosteric activation by TPX2 of dephosphorylated Aurora A could be at play in the spindle microtubules. The mechanistic insights into autophosphorylation and allosteric activation by TPX2 binding proposed here, may have implications for understanding regulation of other protein kinases.

scholarly journals Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens

10.3791/54991 ◽  
2017 ◽  
Author(s):  
Bruce J. MacLachlan ◽  
Alexander Greenshields-Watson ◽  
Georgina H Mason ◽  
Andrea J Schauenburg ◽  
Valentina Bianchi ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
X Ray ◽  
X Ray Crystallography ◽  
Functional Assays ◽  
Cancer Antigens ◽  
Receptor Recognition

scholarly journals General discussion summary

2002 ◽  
Vol 357 (1426) ◽  
pp. 1419-1420 ◽  
Keyword(s):  
Water Splitting ◽  
Molecular Mechanisms ◽  
Structure And Function ◽  
X Ray ◽  
X Ray Crystallography ◽  
And Function ◽  
Splitting Process

This general discussion was chaired by A. W. Rutherford ( Service de Bioénergétique, Saclay, France ) and revolved around two major topics: (i) the implications of X–ray crystallography on the relationships between structure and function; (ii) the molecular mechanisms of the water–splitting process.

scholarly journals Molecular mechanisms of enzyme dysfunction in human phosphoglucomutase-1 deficiency

2019 ◽  
Author(s):  
◽  
Kyle Matthew Stiers
Keyword(s):  
Molecular Mechanisms ◽  
Structural Features ◽  
Therapeutic Interventions ◽  
Structural Studies ◽  
X Ray ◽  
Missense Variants ◽  
X Ray Crystallography ◽  
Ancient Lineage ◽  
Research Opportunity

Human phosphoglucomutase-1 (PGM1) belongs to the [alpha]-D-phosphohexomutase superfamily, an ancient lineage of enzymes critical for carbohydrate metabolism. PGM1 catalyzes the interconversion of glucose-1-phosphate and glucose-6-phosphate, acting as the pivot between energy storage and utilization. Recently, PGM1 has been implicated as the monogenic cause of an inherited metabolic disease in humans, called PGM1 deficiency. The disease presents with highly variable phenotype in patients and is difficult to diagnose. Furthermore, genotype-phenotype relationships remain unclear-even in siblings with the same missense variants, no obvious correlation exists. PGM1 deficiency is a unique research opportunity due to the lack of clear rationale for varying effects of missense variants, availability of patient data, favorable in vitro behavior of recombinantly expressed PGM1, and the limited number of structural studies characterizing individual missense variants. In this work we have characterized multiple molecular mechanisms of disease through X-ray crystallography and biochemistry. Thus, this work provides a foundation for physicians to make much more accurate prognostic decisions when advising patients, identifies variants with possible therapeutic interventions, and informs us of key dynamics and structural features required for proper functioning of human PGM1.

scholarly journals Author response: Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2

2014 ◽  
Author(s):  
Adelajda Zorba ◽  
Vanessa Buosi ◽  
Steffen Kutter ◽  
Nadja Kern ◽  
Francesco Pontiggia ◽  
...  
Keyword(s):  
Molecular Mechanism ◽  
Author Response ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Allosteric Activation

scholarly journals Allosteric Regulation of the EphA2 Receptor Intracellular Region by Serine/Threonine Kinases

2021 ◽  
Author(s):  
Bernhard C. Lechtenberg ◽  
Marina P. Gehring ◽  
Taylor P. Light ◽  
Mike W. Matsumoto ◽  
Kalina Hristova ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Molecular Mechanisms ◽  
Structural Information ◽  
Cell Communication ◽  
Kinase Domain ◽  
Eph Receptor ◽  
X Ray ◽  
X Ray Crystallography ◽  
Exchange Mass Spectrometry ◽  
Intracellular Region

ABSTRACTEph receptor tyrosine kinases play a key role in cell-cell communication. However, lack of structural information on the entire multi-domain intracellular region of any Eph receptor has hindered detailed understanding of their signaling mechanisms. Here, we use an integrative structural biology approach combining X-ray crystallography, small-angle X-ray scattering and hydrogen-deuterium exchange mass spectrometry, to gain the first insights into the structure and dynamics of the entire EphA2 intracellular region. EphA2 promotes cancer malignancy through a poorly understood non-canonical form of signaling that depends on serine/threonine phosphorylation of the linker connecting the EphA2 kinase and SAM domains. We uncovered two distinct molecular mechanisms that may function in concert to mediate the effects of linker phosphorylation through an orchestrated allosteric regulatory network. The first involves a shift in the equilibrium between a “closed” configuration of the EphA2 intracellular region and an “open” more extended configuration induced by the accumulation of phosphorylation sites in the linker. This implies that cooperation of multiple serine/threonine kinase signaling networks is necessary to promote robust EphA2 non-canonical signaling. The second involves allosteric rearrangements in the kinase domain and juxtamembrane segment induced by phosphorylation of some linker residues, suggesting a link between EphA2 non-canonical signaling and canonical signaling through tyrosine phosphorylation. Given the key role of EphA2 in cancer malignancy, this new knowledge can inform therapeutic strategies.

scholarly journals A new approach to time-resolved X-ray crystallography

2014 ◽  
Vol 70 (a1) ◽  
pp. C1434-C1434
Author(s):  
Briony Yorke ◽  
Arwen Pearson ◽  
Godfrey Beddard ◽  
Robin Owen
Keyword(s):  
Molecular Mechanisms ◽  
Structural Information ◽  
Information Storage ◽  
High Time Resolution ◽  
Time Resolved ◽  
New Approach ◽  
X Ray ◽  
X Ray Crystallography ◽  
Current State ◽  
Initial Results

Time-resolved crystallography is able to provide four-dimensional structural information about short-lived intermediate states, with near-atomic resolution. This information can be used to elucidate molecular mechanisms relevant to areas such as drug-design, chemical and biological sensors, and energy and information storage. The current state of the art time-resolved experiments can reach picosecond time-resolutions using Laue crystallography but such experiments can only be carried out at a few beamlines worldwide.We have developed a new transform time-resolved method that can be performed using a monochromatic beamline at a synchrotron and still achieve high time-resolution, vastly increasing the accessibility of such experiments. Here we present initial results demonstrating the method.

scholarly journals Structural insights into the small G-protein Arl13B and implications for Joubert syndrome

2013 ◽  
Vol 457 (2) ◽  
pp. 301-311 ◽  
Author(s):  
Mandy Miertzschke ◽  
Carolin Koerner ◽  
Michael Spoerner ◽  
Alfred Wittinghofer
Keyword(s):  
High Resolution ◽  
G Protein ◽  
Joubert Syndrome ◽  
Active Conformation ◽  
Loss Of Function ◽  
X Ray ◽  
X Ray Crystallography ◽  
Small G Protein ◽  
The Stability ◽  
Structural Insights

Using Arl13B from Chlamydomonas as a model, we show by high resolution X-ray crystallography and biochemical approaches that mutations in patients with Joubert syndrome might lead to loss of function by specifically affecting the stability of the active conformation of Arl13B.

Abstract 3902: Development of Aurora A inhibitors withortho-halophenyl substituted pyrimidines: Unusual potency, SAR and X-ray crystallography studies.

2012 ◽  
Author(s):  
Harshani R. Lawrence ◽  
Mathew P. Martin ◽  
Yunting Luo ◽  
Roberta Pireddu ◽  
Harsukh Gevariya ◽  
...  
Keyword(s):  
Aurora A ◽  
X Ray ◽  
X Ray Crystallography
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