The use of conformationally thermostabilised GPCRs in drug discovery: application to fragment, structure and biophysical techniques

Tehan, Benjamin G.; Christopher, John

doi:10.1016/j.coph.2016.06.010

Cited by 13 publications

(4 citation statements)

References 26 publications

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“…Unlike the aMD/MSM ensemble, the resulting aMD ensemble predicts that CypA adopts predominantly a 100sopen/70sopen conformation (Fig. S11) Approaches have been developed based on using antibodies to stabilise transiently populated conformation states to facilitate ligand discovery efforts [79][80][81] . Our results open an exciting opportunity for determining this in silico as part of rational drug design endeavours.…”

Section: Discussionmentioning

confidence: 99%

Dynamic design: manipulation of millisecond timescale motions on the energy landscape of Cyclophilin A

Juárez-Jiménez

Gupta

Karunanithy

et al. 2018

Preprint

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Proteins need to interconvert between many conformations in order to function, many of which are formed transiently, and sparsely populated. Particularly when the lifetimes of these states approach the millisecond timescale, identifying the relevant structures and the mechanism by which they inter-convert remains a tremendous challenge. Here we introduce a novel combination of accelerated MD (aMD) simulations and Markov State modelling (MSM) to explore these 'excited' conformational states . Applying this to the highly dynamic protein CypA, a protein involved in immune response and associated with HIV infection, we identify five principally populated conformational states and the atomistic mechanism by which they interconvert. A rational design strategy predicted that the mutant D66A should stabilise the minor conformations and substantially alter the dynamics whereas the similar mutant H70A should leave the landscape broadly unchanged. These predictions are confirmed using CPMG and R1ρ solution state NMR measurements. By accurately and reliably exploring functionally relevant, but sparsely populated conformations with milli-second lifetimes in silico, our aMD/MSM method has tremendous promise for the design of dynamic protein free energy landscapes for both protein engineering and drug discovery.

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Section: Discussionmentioning

confidence: 99%

Dynamic design: manipulation of millisecond timescale motions on the energy landscape of Cyclophilin A

Juárez-Jiménez

Gupta

Karunanithy

et al. 2018

Preprint

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“…Recent advances in crystallisation techniques for GPCRs (such as thermo-stabilised receptors: StaRs; [26] , [27] ) have facilitated the large-scale purification of stable GPCRs in a conformation appropriate for both X-ray crystallography and their use as antigens for monoclonal antibody generation. Previously, Hutchings et al [28] characterised a series of monoclonal antibodies (mAbs) that were raised against the thermo-stabilised turkey β 1 -adrenoceptor (β 1 AR-m23 StaR) [8] .…”

Section: Introductionmentioning

confidence: 99%

A monoclonal antibody raised against a thermo-stabilised β1-adrenoceptor interacts with extracellular loop 2 and acts as a negative allosteric modulator of a sub-set of β1-adrenoceptors expressed in stable cell lines

Soave

Cseke

Hutchings

et al. 2018

Biochemical Pharmacology

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“…Conversely, cyclic AMP synthesis can be moderated through inhibitory G (G i ) GPCR signals [24,25]. Similarly, cyclic GMP can be generated via activation of membrane-bound guanylate cyclase through peptide hormones (such as atrial natriuretic factor) and/or through intracellular activation of soluble guanylate cyclase (by nitric oxide, carbon monoxide).…”

Section: Precision Medicine For Vsm-directed Cyclic Nucleotide-mediamentioning

confidence: 99%

Novel protein kinase targets in vascular smooth muscle therapeutics

Tulis

2017

Current Opinion in Pharmacology

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Many signaling factors have been identified over the years that serve as mechanistic foundations for the pathogenesis and/or maintenance of cardiovascular disease (CVD). Of these, cyclic nucleotide-driven protein kinases in vascular smooth muscle (VSM) are of essential importance. Comprised primarily of cyclic AMP-dependent and cyclic GMP-dependent protein kinases, these ubiquitous signaling molecules have capacity to operate through numerous downstream effectors including vasodilator-stimulated phosphoprotein (VASP) to control aberrant VSM growth elemental to CVD. As more information is gathered regarding genetic, biochemical, molecular and cellular makeup of CVD including VSM cyclic nucleotide-dependent protein kinases and VASP, advances will be made in precision medicine by identifying more precise therapeutic targets to enhance clinical decision making.

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The use of conformationally thermostabilised GPCRs in drug discovery: application to fragment, structure and biophysical techniques

Cited by 13 publications

References 26 publications

Dynamic design: manipulation of millisecond timescale motions on the energy landscape of Cyclophilin A

Dynamic design: manipulation of millisecond timescale motions on the energy landscape of Cyclophilin A

A monoclonal antibody raised against a thermo-stabilised β1-adrenoceptor interacts with extracellular loop 2 and acts as a negative allosteric modulator of a sub-set of β1-adrenoceptors expressed in stable cell lines

Novel protein kinase targets in vascular smooth muscle therapeutics

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