Paper

Site-specific quantification of the in vivo UFMylome reveals myosin modification in ALS

Lewis, Christopher T. A., Stokes, Matthew P., Ngo, Shyuan T., Zee, Barry M., Ng, Yaan-Kit, Steyn, Frederik J., Peckham, Hayley, McNamara, James W., Gregorevic, Paul, Goodman, Craig A., Ochala, Julien, Zhang, Chengxin, Parker, Benjamin L., Blazev, Ronnie

2024

Overview

UFMylation is a ubiquitin-like protein post-translational modification of Ubiquitin Fold Modifier 1 (UFM1) applied to substrate proteins. The UFMylation system is important for normal development and plays a critical role in a variety of cellular processes including regulating telomere length, stress responses and protein quality control. Here, we describe the development of an antibody-based enrichment approach to immunoprecipitate in vivo remnant UFMylated peptides and identification by liquid chromatography and tandem mass spectrometry (LC-MS/MS). We used this approach to identify >200 UFMylation sites from various mouse tissues revealing extensive modification in skeletal muscle. Furthermore, we show that UFMylation is increased in skeletal muscle biopsies from people living with Amyotrophic Lateral Sclerosis (plwALS). Quantification of UFMylation sites in these participant biopsies with multiplexed isotopic labeling and LC-MS/MS reveal prominent increases in myosin UFMylation. Finally, in silico modelling suggest UFMylation of myosin directly adjacent to the ATP-binding site may regulate stability and/or function. Our data suggest that although UFMylation is not as widespread as ubiquitylation, its in vivo status is more complex than previously thought.

• Publisher: Cold Spring Harbor Laboratory
• Subject: Biochemistry