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• W3127002146 abstract "Abstract We describe the crystal structure determination of the ARM domain of Drosophila SARM1 (dSARM1 ARM ), which required combination of a number of sources of phase information in order to obtain interpretable electron density maps. SARM1 is a central executioner of the process of axon degeneration, a common feature of the early phase of a range of neurodegenerative diseases. SARM1 is held in the inactive state in healthy axons by its N-terminal auto-inhibitory ARM domain, and is activated to cleave NAD + upon injury, triggering the subsequent axon degeneration. To characterize the molecular mechanism of SARM1 activation, we sought to determine the crystal structure of the SARM1 ARM domain. Here we describe the recombinant production and crystallization of dSARM1 ARM , as well as unconventional process used for structure determination. Crystals were obtained in the presence of NMN, a precursor of NAD + and a potential activator of SARM1, only after in situ proteolysis of the N-terminal 63 residues. After molecular replacement attempts failed, we determined the crystal structure of dSARM1 ARM at 1.65 Å resolution using the MIRAS phasing technique with the program autoSHARP, combining data from the native, SeMet-labelled, and Br-soaked crystals. The structure will further our understanding of the regulation of SARM1." @default.
• W3127002146 created "2021-02-15" @default.
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• W3127002146 date "2021-02-01" @default.
• W3127002146 modified "2023-10-16" @default.
• W3127002146 title "Crystal structure determination of the armadillo repeat domain of Drosophila SARM1 using MIRAS phasing" @default.
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• W3127002146 doi "https://doi.org/10.1101/2021.01.31.428505" @default.
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