Secondary Lipid Oxidation Products as Modulators of Calpain-2 Functionality In Vitro

ABSTRACT: Secondary lipid oxidation products modify and impact the function of proteins. Identifying where and how these products modify a protein's structure would allow for targeted approaches to mitigate or prevent this modification. Here, we purified calpain-2, a neutral calcium dependent cysteine protease, from porcine skeletal muscle and conducted in-vitro experiments using various concentrations of malondialdehyde, 2-hexenal, and 4-hydroxynonenal. We determined these secondary lipid oxidation products differentially impact calpain-2 proteolytic activity and autolysis. We identified modifications of malondialdehyde and hexenal on calpain-2 with LC-MS/MS approaches. We further evaluated the number of adduction sites on the catalytic and regulatory subunits of calpain-2 through MALDI-MS approaches. These data demonstrate that secondary lipid oxidation products can modify and differentially impact calpain-2.

INSTRUMENT(S): Orbitrap Eclipse, ultrafleXtreme

ORGANISM(S): Sus Scrofa (ncbitaxon:9823)

SUBMITTER: Steven Lonergan  

PROVIDER: MSV000093040 | MassIVE | Wed Oct 04 15:10:00 BST 2023

SECONDARY ACCESSION(S): PXD045905

REPOSITORIES: MassIVE