University of Massachusetts Medical School Faculty Publications

Title

Initial Kinetic Characterization of Sterile Alpha and Toll/Interleukin Receptor Motif-Containing Protein 1

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology; Program in Chemical Biology; Graduate School of Biomedical Sciences; Thompson Lab

Publication Date

2020-02-12

Document Type

Article

Disciplines

Amino Acids, Peptides, and Proteins | Biochemistry | Enzymes and Coenzymes | Medicinal-Pharmaceutical Chemistry | Molecular and Cellular Neuroscience | Nervous System Diseases | Nucleic Acids, Nucleotides, and Nucleosides

Abstract

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.

Keywords

Peptides and proteins, Assays, Inhibitors, Inhibition, Nicotinamide

DOI of Published Version

10.1021/acs.biochem.9b01078

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

Related Resources

Link to Article in PubMed

Journal/Book/Conference Title

Biochemistry

PubMed ID

32049506

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