Browsing by Author "Anowai, Anthonia"

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    N-terminomics/proteomics investigation of Calpain-2 substrates
    (2022-04-21) Anowai, Anthonia; Dufour, Antoine; Hollenberg, Morley; Robbins, Stephen
    Calpains are a family of intracellular cysteine proteases that require calcium for activation. Calpain-1 and -2, first described as the classical calpains due to their ubiquitous expression, are part of this larger 15-member family in humans and share similar patterns of expression and structural domains. Despite suggestive indication of a role for calpains in different cell functions in disease and health, it is unclear the repertoire of substrates that are targeted by calpains 1 and 2 in macrophages. To investigate this hypothesis, we focused on calpain-2 and performed N-terminomics/TAILS experiment in calpain-2 knockout human THP-1 monocytic cell lines to identify new substrates. We analyzed the proteomics and Nterminomics data using bioinformatics and pathway enrichment tools (STRING-db (https://string-db.org) and Metascape (https://metascape.org/gp/index.html)). We validated proteolytically processed substrates of calpain-2. We have shown that calpain-2 has novel targets spleen tyrosine kinase (SYK) in THP-1 cells when activated by calcium and may play a role in regulating kinase.