Structural Analysis of SOD1 Fibrils with Mass Spectrometry, Limited Proteolysis, and Atomic Force Microscopy (AFM)

Bon-Kyung Koo; Julian Whitelegge

doi:10.1007/978-1-0716-2597-2_30

Structural Analysis of SOD1 Fibrils with Mass Spectrometry, Limited Proteolysis, and Atomic Force Microscopy (AFM)

Methods Mol Biol. 2023:2551:481-495. doi: 10.1007/978-1-0716-2597-2_30.

Authors

Bon-Kyung Koo¹, Julian Whitelegge²

Affiliations

¹ The Department of Chemistry and Biochemistry, School of Physical Sciences, University of California, Los Angeles, USA.
² The Pasarow Mass Spectrometry Laboratory, The Jane and Terry Semel Institute for Neuroscience and Human Behavior, David Geffen School of Medicine, University of California, Los Angeles, USA. jpw@chem.ucla.edu.

PMID: 36310221
DOI: 10.1007/978-1-0716-2597-2_30

Abstract

This protocol describes a method to purify SOD1 in Saccharomyces cerevisiae to characterize using ICP-MS and AFM, to agitate and fibrillate for aggregation of SOD1. The human SOD1 (hSOD1) is a 32-kDa homodimer, with one copper- and one zinc-binding site per 153-amino acid subunit. Misfolded protein aggregates are often correlated with diseases known as amyloidosis, including ALS, Alzheimer's, Parkinson's, and prion disease (Valentine and Hart, Proc Natl Acad Sci USA 100: 3617-3622, 2003; Tanzi and Bertram, Cell 120: 545-555, 2005; Soto and Pritzkow, Nat Neurosci 21:1332-1340, 2018; Sarafian et al., J Neurosci Res 95:1871-1887, 2017). Proteinaceous aggregates containing hSOD1 have frequently been found in the spinal cords of ALS patients.

Keywords: AFM; ALS (Amyotrophic lateral sclerosis); Aggregation; Cu/Zn SOD1 (Superoxide dismutase1); Fibrillation; ICP-MS.

MeSH terms

Amyotrophic Lateral Sclerosis*
Humans
Mass Spectrometry
Microscopy, Atomic Force
Mutation
Protein Aggregates
Proteolysis
Superoxide Dismutase / metabolism
Superoxide Dismutase-1 / genetics
Superoxide Dismutase-1 / metabolism

Substances

Superoxide Dismutase-1
Superoxide Dismutase
Protein Aggregates
SOD1 protein, human