Initial Kinetic Characterization of Sterile Alpha and Toll/Interleukin Receptor Motif-Containing Protein 1
UMass Chan Affiliations
Thompson LabGraduate School of Biomedical Sciences
Program in Chemical Biology
Department of Biochemistry and Molecular Pharmacology
Document Type
Journal ArticlePublication Date
2020-02-12Keywords
Peptides and proteinsAssays
Inhibitors
Inhibition
Nicotinamide
Amino Acids, Peptides, and Proteins
Biochemistry
Enzymes and Coenzymes
Medicinal-Pharmaceutical Chemistry
Molecular and Cellular Neuroscience
Nervous System Diseases
Nucleic Acids, Nucleotides, and Nucleosides
Metadata
Show full item recordAbstract
Sterile alpha and toll/interleukin receptor (TIR) motif-containing protein 1 (SARM1) plays a pivotal role in triggering the neurodegenerative processes that underlie peripheral neuropathies, traumatic brain injury, and neurodegenerative diseases. Importantly, SARM1 knockdown or knockout prevents degeneration, thereby demonstrating that SARM1 is a promising therapeutic target. Recently, SARM1 was shown to promote neurodegeneration via its ability to hydrolyze NAD(+), forming nicotinamide and ADP ribose (ADPR). Herein, we describe the initial kinetic characterization of full-length SARM1, as well as the truncated constructs corresponding to the SAM(1-2)TIR and TIR domains, highlighting the distinct challenges that have complicated efforts to characterize this enzyme. Moreover, we show that bacterially expressed full-length SARM1 (kcat/KM = 6000 +/- 2000 M(-1) s(-1)) is at least as active as the TIR domain alone (kcat/KM = 1500 +/- 300 M(-1) s(-1)). Finally, we show that the SARM1 hydrolyzes NAD(+) via an ordered uni-bi reaction in which nicotinamide is released prior to ADPR.Source
Loring HS, Icso JD, Nemmara VV, Thompson PR. Initial Kinetic Characterization of Sterile Alpha and Toll/Interleukin Receptor Motif-Containing Protein 1. Biochemistry. 2020 Mar 3;59(8):933-942. doi: 10.1021/acs.biochem.9b01078. Epub 2020 Feb 17. PMID: 32049506. Link to article on publisher's site
DOI
10.1021/acs.biochem.9b01078Permanent Link to this Item
http://hdl.handle.net/20.500.14038/29438PubMed ID
32049506Related Resources
ae974a485f413a2113503eed53cd6c53
10.1021/acs.biochem.9b01078