Engineering of a protein probe with multiple inputs and multiple outputs for evaluation of alpha synuclein aggregation states

The aggregation of α-synuclein (αS) into oligomers and fibrils is implicated in the pathology of Parkinson's Disease (PD). While a molecular probe for rapid and comprehensive evaluation of αS aggregation states is critical for a better understanding of PD pathology, identification of therapeuti...

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Publié dans:Biochemical engineering journal. - 1998. - 178(2022) vom: 04. Jan.
Auteur principal: Chau, Edward (Auteur)
Autres auteurs: Kim, Jin Ryoun
Format: Article en ligne
Langue:English
Publié: 2022
Accès à la collection:Biochemical engineering journal
Sujets:Journal Article Aggregation Alpha-synuclein Amyloid Fibril Oligomer Protein probe
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520 |a The aggregation of α-synuclein (αS) into oligomers and fibrils is implicated in the pathology of Parkinson's Disease (PD). While a molecular probe for rapid and comprehensive evaluation of αS aggregation states is critical for a better understanding of PD pathology, identification of therapeutic candidates, and the development of early diagnostic strategies, no such probe has yet to be developed. A structurally flexible αS variant, PG65, was previously developed as a target binding-driven, conformation-switching molecular probe for rapid αS oligomer detection. Though informative, detection using PG65 provides no comprehensive assessment of the αS aggregation states. In the present study, we report engineering of a molecular probe, PG65-MIMO (a PG65 variant with Multiple-Inputs and Multiple-Outputs), that rapidly (within 2 hr) produces comprehensive information on αS aggregation states. PG65-MIMO generates distinct fluorescence responses to the three major αS conformers (monomers, oligomers, and fibrils). PG65-MIMO also displays unique fluorescent signals for αS oligomers, depending on the tris(2-carboxyethyl)phosphine (TCEP) concentration. Our results suggest that the TCEP dependent signaling of PG65-MIMO may be associated with its conformational states. Overall, our study illustrates engineering of an αS variant to create a molecular probe for handling multiple inputs and multiple outputs, addressing the technological gap in αS detection 
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700 1 |a Kim, Jin Ryoun  |e verfasserin  |4 aut 
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