Deep mutational scanning reveals the structural basis for α-synuclein activity.

Nature Chemical Biology
Robert W NewberryWilliam F DeGrado

Abstract

Defining the biologically active structures of proteins in their cellular environments remains challenging for proteins with multiple conformations and functions, where only a minor conformer might be associated with a given function. Here, we use deep mutational scanning to probe the structure and dynamics of α-synuclein, a protein known to adopt disordered, helical and amyloid conformations. We examined the effects of 2,600 single-residue substitutions on the ability of intracellularly expressed α-synuclein to slow the growth of yeast. Computational analysis of the data showed that the conformation responsible for this phenotype is a long, uninterrupted, amphiphilic helix with increasing dynamics toward the C terminus. Deep mutational scanning can therefore determine biologically active conformations in cellular environments, even for a highly dynamic multi-conformational protein.

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Citations

Jan 27, 2021·Annual Review of Pathology·Gautam Runwal, Robert H Edwards
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Oct 26, 2021·Faraday Discussions·Huong T KratochvilWilliam F DeGrado

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Methods Mentioned

BETA
X-ray
electron microscopy
electron paramagnetic resonance
NMR
circular dichroism
circular
fluorescence microscopy
Illumina sequencing
PCR
dynamic light scattering

Software Mentioned

Bowtie2
TANGO
NIS
Elements

Related Concepts

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Alpha-Synuclein Aggregation (MDS)

Alpha-synucleins are small proteins that are believed to restrict the mobility of synpatic vesicles and inhibit neurotransmitter release. Aggregation of these proteins have been linked to several types of neurodegenerative diseases including dementia with Lewy bodies and Parkinson's disease. Here is the latest research on α-synuclein aggregation.

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