TY - JOUR
T1 - Identification of TrkB Binders from Complex Matrices Using a Magnetic Drug Screening Nanoplatform
AU - Arituluk, Zekiye Ceren
AU - Horne, Jesse
AU - Adhikari, Bishnu
AU - Steltzner, Jeffrey
AU - Mansur, Shomit
AU - Ahirwar, Parmanand
AU - Velu, Sadanandan E.
AU - Gray, Nora E.
AU - Ciesla, Lukasz M.
AU - Bao, Yuping
N1 - Funding Information:
This work was supported by NSF-CBET 1915873 and Merrymac-McKinley Foundation Award. Acknowledgments
Funding Information:
This work was supported by NSF-CBET 1915873 and Merrymac-McKcinley Foundation Award. Y. B. and L.M.C. acknowledge the Alabama Life Research Institute for seed funding support and the University of Alabama Department of Biological Sciences for the use of TEM and confocal microscopy.
Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/8/16
Y1 - 2021/8/16
N2 - Brain-derived neurotrophic factor (BDNF) and its receptor tyrosine receptor kinase B (TrkB) have been shown to play an important role in numerous neurological disorders, such as Alzheimer's disease. The identification of biologically active compounds interacting with TrkB serves as a drug discovery strategy to identify drug leads for neurological disorders. Here, we report effective immobilization of functional TrkB on magnetic iron oxide nanoclusters, where TrkB receptors behave as "smart baits"to bind compounds from mixtures and magnetic nanoclusters enable rapid isolation through magnetic separation. The presence of the immobilized TrkB was confirmed by specific antibody labeling. Subsequently, the activity of the TrkB on iron oxide nanoclusters was evaluated with ATP/ADP conversion experiments using a known TrkB agonist. The immobilized TrkB receptors can effectively identify binders from mixtures containing known binders, synthetic small molecule mixtures, and Gotu Kola (Centella asiatica) plant extracts. The identified compounds were analyzed by an ultrahigh-performance liquid chromatography system coupled with a quadrupole time-of-flight mass spectrometer. Importantly, some of the identified TrkB binders from Gotu Kola plant extracts matched with compounds previously linked to neuroprotective effects observed for a Gotu Kola extract approved for use in a clinical trial. Our studies suggest that the possible therapeutic effects of the Gotu Kola plant extract in dementia treatment, at least partially, might be associated with compounds interacting with TrkB. The unique feature of this approach is its ability to fast screen potential drug leads using less explored transmembrane targets. This platform works as a drug-screening funnel at early stages of the drug discovery pipeline. Therefore, our approach will not only greatly benefit drug discovery processes using transmembrane proteins as targets but also allow for evaluation and validation of cellular pathways targeted by drug leads.
AB - Brain-derived neurotrophic factor (BDNF) and its receptor tyrosine receptor kinase B (TrkB) have been shown to play an important role in numerous neurological disorders, such as Alzheimer's disease. The identification of biologically active compounds interacting with TrkB serves as a drug discovery strategy to identify drug leads for neurological disorders. Here, we report effective immobilization of functional TrkB on magnetic iron oxide nanoclusters, where TrkB receptors behave as "smart baits"to bind compounds from mixtures and magnetic nanoclusters enable rapid isolation through magnetic separation. The presence of the immobilized TrkB was confirmed by specific antibody labeling. Subsequently, the activity of the TrkB on iron oxide nanoclusters was evaluated with ATP/ADP conversion experiments using a known TrkB agonist. The immobilized TrkB receptors can effectively identify binders from mixtures containing known binders, synthetic small molecule mixtures, and Gotu Kola (Centella asiatica) plant extracts. The identified compounds were analyzed by an ultrahigh-performance liquid chromatography system coupled with a quadrupole time-of-flight mass spectrometer. Importantly, some of the identified TrkB binders from Gotu Kola plant extracts matched with compounds previously linked to neuroprotective effects observed for a Gotu Kola extract approved for use in a clinical trial. Our studies suggest that the possible therapeutic effects of the Gotu Kola plant extract in dementia treatment, at least partially, might be associated with compounds interacting with TrkB. The unique feature of this approach is its ability to fast screen potential drug leads using less explored transmembrane targets. This platform works as a drug-screening funnel at early stages of the drug discovery pipeline. Therefore, our approach will not only greatly benefit drug discovery processes using transmembrane proteins as targets but also allow for evaluation and validation of cellular pathways targeted by drug leads.
KW - BDNF/TrkB
KW - drug discovery
KW - magnetic nanoclusters
KW - neurologic disorders
KW - plant extract
KW - receptor immobilization
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U2 - 10.1021/acsabm.1c00552
DO - 10.1021/acsabm.1c00552
M3 - Article
C2 - 35006910
AN - SCOPUS:85112565482
SN - 2576-6422
VL - 4
SP - 6244
EP - 6255
JO - ACS Applied Bio Materials
JF - ACS Applied Bio Materials
IS - 8
ER -