N-terminomics/proteomics investigation of Calpain-2 substrates

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.

Name thesis:degree_name

Master of Science (MSc)

Discipline thesis:degree_discipline

Medicine – Biochemistry and Molecular Biology

Grantor dc:publisher.institution

Science

Year dc:date.issued

2022

Author dc:creator

• Anowai, Anthonia

Advisor dc:contributor.advisor

• Dufour, Antoine

Committee members dc:contributor.committeemember

• Hollenberg, Morley
• Robbins, Stephen

• proteases proteomics

Statement dc:rights

• University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.

Language dc:language.iso

eng

DOI

http://dx.doi.org/10.11575/PRISM/39702

OAI identifier oai:identifier

oai:ucalgary.scholaris.ca:1880/114581