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Identification of Small Molecule Inhibitors of Tau Aggregation by Targeting Monomeric Tau As a Potential Therapeutic Approach for Tauopathies

View Article: PubMed Central

ABSTRACT

A potential strategy to alleviate the aggregation of intrinsically disordered proteins (IDPs) is to maintain the native functional state of the protein by small molecule binding. However, the targeting of the native state of IDPs by small molecules has been challenging due to their heterogeneous conformational ensembles. To tackle this challenge, we applied a high-throughput chemical microarray surface plasmon resonance imaging screen to detect the binding between small molecules and monomeric full-length Tau, a protein linked with the onset of a range of Tauopathies. The screen identified a novel set of drug-like fragment and lead-like compounds that bound to Tau. We verified that the majority of these hit compounds reduced the aggregation of different Tau constructs in vitro and in N2a cells. These results demonstrate that Tau is a viable receptor of drug-like small molecules. The drug discovery approach that we present can be applied to other IDPs linked to other misfolding diseases such as Alzheimer’s and Parkinson’s diseases.

No MeSH data available.


Related in: MedlinePlus

The HT-CM-SPR scheme. (A) The protein analyte is allowed to float over the array surface under controlled conditions to allow binding events to happen. SPR Imaging enables the detection of binding events: (B) Close-up of a grey scale picture obtained by CCD camera imaging of chemical microarray. (C) Grey scale analysis resulted in the parallel detection of 9,216 SPR minima per microarray exhibiting a shift in the resonance wavelength upon protein binding to the immobilized compounds. (D) Generic example of a color coded visualization of one array experiment.
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Figure 1: The HT-CM-SPR scheme. (A) The protein analyte is allowed to float over the array surface under controlled conditions to allow binding events to happen. SPR Imaging enables the detection of binding events: (B) Close-up of a grey scale picture obtained by CCD camera imaging of chemical microarray. (C) Grey scale analysis resulted in the parallel detection of 9,216 SPR minima per microarray exhibiting a shift in the resonance wavelength upon protein binding to the immobilized compounds. (D) Generic example of a color coded visualization of one array experiment.

Mentions: In this study, we pursue an alternative approach to those reported previously for identifying small molecule inhibitors of Tau [7-11]. First, we apply a high-throughput chemical microarray surface plasmon resonance imaging screen (HT-CM-SPR) [28, 34-36], which has the ability to detect the interaction between immobilized small molecules and monomeric full length Tau (hTau2N4Rwt), to probe whether the binding of fragments and/or lead-like compounds to Tau is feasible. In contrast to more common SPR approaches, in which the protein target itself is immobilized on the sensor surface, this “reverse SPR” scheme exploits the advantages of having large collections of compounds immobilized on chemical microarrays (Fig. 1). Because the surface chemistry applied for the chemical microarray allows controlling the orientation and density of the compounds precisely, the technique is applicable to the screening of a variety of biomolecules (including structured and intrinsically disorder proteins, and antibodies) with a variety of structures. This universal screening platform has been a powerful tool for the identification of small molecule binders to proteins providing valuable starting points for hit-to-lead optimization in various drug discovery projects [28, 34-36]. Next, we identified novel small molecule binders capable of modulating Tau aggregation in vitro and in N2a cells. Our overall goal is to identify drug-like small molecules that bind to monomeric Tau and can reduce its aggregation. By specifically targeting the monomeric state of Tau, we anticipate that it will be possible to identify small molecules that modulate Tau aggregation at the earliest phases of its fibrillization pathway.


Identification of Small Molecule Inhibitors of Tau Aggregation by Targeting Monomeric Tau As a Potential Therapeutic Approach for Tauopathies
The HT-CM-SPR scheme. (A) The protein analyte is allowed to float over the array surface under controlled conditions to allow binding events to happen. SPR Imaging enables the detection of binding events: (B) Close-up of a grey scale picture obtained by CCD camera imaging of chemical microarray. (C) Grey scale analysis resulted in the parallel detection of 9,216 SPR minima per microarray exhibiting a shift in the resonance wavelength upon protein binding to the immobilized compounds. (D) Generic example of a color coded visualization of one array experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4997948&req=5

Figure 1: The HT-CM-SPR scheme. (A) The protein analyte is allowed to float over the array surface under controlled conditions to allow binding events to happen. SPR Imaging enables the detection of binding events: (B) Close-up of a grey scale picture obtained by CCD camera imaging of chemical microarray. (C) Grey scale analysis resulted in the parallel detection of 9,216 SPR minima per microarray exhibiting a shift in the resonance wavelength upon protein binding to the immobilized compounds. (D) Generic example of a color coded visualization of one array experiment.
Mentions: In this study, we pursue an alternative approach to those reported previously for identifying small molecule inhibitors of Tau [7-11]. First, we apply a high-throughput chemical microarray surface plasmon resonance imaging screen (HT-CM-SPR) [28, 34-36], which has the ability to detect the interaction between immobilized small molecules and monomeric full length Tau (hTau2N4Rwt), to probe whether the binding of fragments and/or lead-like compounds to Tau is feasible. In contrast to more common SPR approaches, in which the protein target itself is immobilized on the sensor surface, this “reverse SPR” scheme exploits the advantages of having large collections of compounds immobilized on chemical microarrays (Fig. 1). Because the surface chemistry applied for the chemical microarray allows controlling the orientation and density of the compounds precisely, the technique is applicable to the screening of a variety of biomolecules (including structured and intrinsically disorder proteins, and antibodies) with a variety of structures. This universal screening platform has been a powerful tool for the identification of small molecule binders to proteins providing valuable starting points for hit-to-lead optimization in various drug discovery projects [28, 34-36]. Next, we identified novel small molecule binders capable of modulating Tau aggregation in vitro and in N2a cells. Our overall goal is to identify drug-like small molecules that bind to monomeric Tau and can reduce its aggregation. By specifically targeting the monomeric state of Tau, we anticipate that it will be possible to identify small molecules that modulate Tau aggregation at the earliest phases of its fibrillization pathway.

View Article: PubMed Central

ABSTRACT

A potential strategy to alleviate the aggregation of intrinsically disordered proteins (IDPs) is to maintain the native functional state of the protein by small molecule binding. However, the targeting of the native state of IDPs by small molecules has been challenging due to their heterogeneous conformational ensembles. To tackle this challenge, we applied a high-throughput chemical microarray surface plasmon resonance imaging screen to detect the binding between small molecules and monomeric full-length Tau, a protein linked with the onset of a range of Tauopathies. The screen identified a novel set of drug-like fragment and lead-like compounds that bound to Tau. We verified that the majority of these hit compounds reduced the aggregation of different Tau constructs in vitro and in N2a cells. These results demonstrate that Tau is a viable receptor of drug-like small molecules. The drug discovery approach that we present can be applied to other IDPs linked to other misfolding diseases such as Alzheimer’s and Parkinson’s diseases.

No MeSH data available.


Related in: MedlinePlus